BCMA-targeted CAR-T cell therapy for multiple myeloma

Abstract

This specification provides a method for treating subjects with multiple myeloma who have received one to three prior treatments. The subjects are administered an infusion of chimeric antigen receptor (CAR)-T cells containing CARs that have the ability to specifically bind to the BCMA epitope.

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims priority to U.S. Provisional Patent Application No. 63 / 497,185 filed on 19 April 2023, U.S. Provisional Patent Application No. 63 / 504,184 filed on 24 May 2023, and International Patent Application PCT / US2023 / 031673 filed on 31 August 2023, the disclosures of each of these applications incorporated herein by reference in their entirety.

[0002] Sequence List This application includes a computer-readable sequence listing file submitted with this application in XML format, the contents of which are incorporated herein by reference in their entirety. The sequence listing XML file submitted with this application is titled "14651-063-228_SEQ_LISTING.xml", was created on 24 August 2023, and has a size of 28,450 bytes. [Background technology]

[0003] Multiple myeloma (MSM) is a highly malignant plasma cell neoplasm. It is thought to be a B-cell neoplasm that proliferates uncontrollably in the bone marrow. Symptoms include one or more of the following: hypercalcemia, renal failure, anemia, bone lesions, bacterial infections, hyperviscosity, and amyloidosis. Despite significant improvements in patient outcomes due to the availability of novel therapies, including proteasome inhibitors, immunomodulators, and monoclonal antibodies, MMS remains largely incurable. Many patients either relapse or become resistant to treatment, highlighting the constant need for new therapies for MMS. In particular, lenalidomide-resistant MMS patients who have received 1-3 prior lines of treatment have poor outcomes and progress rapidly through available therapies, highlighting the need for novel, safe, and effective treatment regimens for use in these early-line settings. [Overview of the Initiative] [Means for solving the problem]

[0004] In one embodiment, the Specified Publication provides a method for treating a subject, comprising administering to the subject a certain dose of chimeric antigen receptor (CAR)-containing T cells comprising (a) an extracellular antigen-binding domain having the ability to specifically bind to an epitope of B cell maturation antigen (BCMA), (b) a transmembrane domain, and (c) an intracellular signaling domain. In some embodiments, the subject has multiple myeloma, has received one to three prior lines of treatment, including immunomodulatory drug (IMiD) therapy, and is resistant to IMiD. In some embodiments, the subject has high-risk features, which are optionally cytogenetic abnormalities, International Staging Classification (ISS) stage III, and / or soft tissue plasmacytoma.

[0005] In one embodiment, the Specified Provision provides a method for selectively treating a subject, comprising: (1) determining whether the subject has high-risk features, wherein the high-risk features are cytogenetic abnormalities, International Staging (ISS) stage III, and / or soft tissue plasmacytoma; and (2) administering to a subject determined to have high-risk features in step (1) a certain dose of chimeric antigen receptor (CAR)-containing T cells comprising (a) an extracellular antigen-binding domain having the ability to specifically bind to an epitope of BCMA, (b) a transmembrane domain, and (c) an intracellular signaling domain. In some embodiments, the subject has multiple myeloma, has received one to three prior treatment lines, including IMiD therapy, and is resistant to IMiD.

[0006] In one embodiment, the Specified Information provides a method for selectively treating a subject, comprising administering a certain dose of chimeric antigen receptor (CAR)-containing T cells to a subject determined to have high-risk features, the T cells comprising (a) an extracellular antigen-binding domain having the ability to specifically bind to a BCMA epitope, (b) a transmembrane domain, and (c) an intracellular signaling domain. In some embodiments, the high-risk features are cytogenetic abnormalities, International Staging Classification (ISS) stage III, and / or soft tissue plasmacytoma. In some embodiments, the subject has multiple myeloma, has received one to three prior treatment lines, including IMiD therapy, and is resistant to IMiD.

[0007] In some embodiments of the various methods or aspects provided herein, IMiD is lenalidomide.

[0008] In some embodiments of the various methods or embodiments provided herein, the high-risk feature is a cytogenetic abnormality. In some embodiments, the cytogenetic abnormality is a high-risk cytogenetic abnormality. In some embodiments, the subject has one or more high-risk cytogenetic abnormalities selected from the group including Gain / amp(1q), del(17p), t(4;14), t(14;16), or any combination thereof. In some embodiments, the cytogenetic abnormality includes Gain / amp(1q). In some embodiments, the cytogenetic abnormality includes del(17p). In some embodiments, the cytogenetic abnormality includes t(4;14). In some embodiments, the cytogenetic abnormality includes t(14;16). In some embodiments, the subject has at least two cytogenetic abnormalities. In some embodiments, the subject has at least three cytogenetic abnormalities. In some embodiments, the subject has at least four cytogenetic abnormalities. In some embodiments, the subject has at least five cytogenetic abnormalities. In some embodiments, the subject has at least six cytogenetic abnormalities. In some embodiments, the subject has at least seven cytogenetic abnormalities. In other embodiments, the cytogenetic abnormalities are standard-risk cytogenetic abnormalities. In other embodiments, the high-risk feature is International Staging Classification (ISS) stage III. In yet another embodiment, the high-risk feature is soft tissue plasmacytoma.

[0009] In some embodiments of the various methods or embodiments provided herein, the subject has received one prior treatment line. In some embodiments, the subject has received two prior treatment lines. In some embodiments, the subject has received three prior treatment lines.

[0010] In some embodiments of the various methods or embodiments provided herein, the prior treatment line, or one of the prior treatment lines, includes an IMiD. In some embodiments, the IMiD is or includes pomalidomide. In some embodiments, the IMiD is or includes lenalidomide. In some embodiments, the subject has previously received prior treatment including a combination of lenalidomide and pomalidomide.

[0011] In some embodiments of the various methods or embodiments provided herein, the prior treatment line, or one of the prior treatment lines, comprises an anti-CD38 antibody. In some embodiments, the anti-CD38 antibody is daratumumab and / or isatuximab.

[0012] In some embodiments of the various methods or embodiments provided herein, the pre-treatment line, or one of the pre-treatment lines, comprises a proteasome inhibitor. In some embodiments, the proteasome inhibitor is bortezomib, carfilzomib, ixazomib, or any combination thereof.

[0013] In some embodiments of the various methods or embodiments provided herein, the subject has received further bridging therapy. In certain embodiments, bridging therapy is at the physician's discretion. In some embodiments, bridging therapy comprises pomalidomide, bortezomib, dexamethasone, daratumumab, or any combination thereof. In some embodiments, bridging therapy comprises pomalidomide, bortezomib, and dexamethasone. In other embodiments, bridging therapy comprises daratumumab, pomalidomide, and dexamethasone. In some embodiments, the subject has received bridging therapy approximately every 20 to 30 days. In some embodiments, the subject has received bridging therapy approximately every 21 days. In some embodiments, the subject has received bridging therapy approximately every 28 days. In some embodiments, the subject has received at least 1, 2, 3, 4 cycles, or more of bridging therapy.

[0014] In some embodiments of the various methods or embodiments provided herein, the subject has further received lymphocyte depletion therapy. In some embodiments, lymphocyte depletion therapy comprises daily cyclophosphamide and / or fludarabine. In some embodiments, lymphocyte depletion therapy comprises daily cyclophosphamide and fludarabine. In some embodiments, lymphocyte depletion therapy comprises approximately 300 mg / m² daily for 3 days. 2 The concentration of cyclophosphamide is approximately 30 mg / m². 2 Contains fludarabine at this concentration.

[0015] In some embodiments of the various methods or aspects provided herein, the dose of T cells is 0.5 to 1.0 × 10 6 The cell-to-subject weight is 1 kg. In a preferred embodiment, the T cell dose is approximately 0.75 × 10⁶ 6 The cell-to-subject weight is 1 kg. In some embodiments, the method includes administering a dose of T cells approximately 5 to 7 days after the initiation of lymphocyte depletion therapy. Preferably, the dose is administered as a single infusion.

[0016] In some embodiments of the various methods or aspects provided herein, the method is effective in achieving a total response in a subject after administering a dose of T cells to the subject. In some embodiments, the method is effective in achieving a total response rate of about 70% to about 100%. In some embodiments, the method is effective in achieving a total response rate of about 74%. In some embodiments, the method is effective in achieving a total response rate of about 84.6%. In some embodiments, the method is effective in achieving a total response rate of about 99.4%.

[0017] In some embodiments of the various methods or embodiments provided herein, total response includes, in order from best to worst, (1) severe complete response; (2) complete response; (3) best partial response; (4) partial response; or (5) minimal response. In some embodiments, total response is severe complete response. In some embodiments, the method is effective in achieving severe complete response at rates of about 40% to about 90%, about 50% to about 80%, about 58.2%, or about 68.8%. In other embodiments, total response is complete response. In some embodiments, the method is effective in achieving complete response at rates of about 10% to about 20%. In some embodiments, the method is effective in achieving complete response at rates of about 14.9% or about 17.6%. In some embodiments, total response is best partial response or partial response. In some embodiments, this method is effective in achieving strict complete response or complete response in about 70% to about 90% of cases. In some embodiments, this method is effective in achieving strict complete response or complete response in about 73.1% or about 86.4% of cases. In some embodiments, this method is effective in achieving strict complete response, complete response, or best partial response in about 80% to about 100% of cases. In some embodiments, this method is effective in achieving strict complete response, complete response, or best partial response in about 81.3% or about 96.0% of cases. In some embodiments, this method is effective in achieving minimal response. In some embodiments, this method is effective in further achieving minimal residual disease negativity. In some embodiments, this method is effective in achieving minimal residual disease negativity in about 50% to about 80% of cases. In some embodiments, this method is effective in achieving minimal residual disease negativity in about 60.6% of cases. In some embodiments, this method is effective in achieving minimal residual disease negativity in approximately 71.6% of cases.

[0018] In some embodiments of the various methods or aspects provided herein, the method is effective in helping at least about 60 to about 100% of subjects to achieve further 12-month progression-free survival. In some embodiments, the method is effective in helping at least about 69.4 to about 81.1% of subjects to achieve further 12-month progression-free survival. In some embodiments, the method is effective in helping at least about 84.1 to about 93.4% of subjects to achieve further 12-month progression-free survival. In some embodiments, the method is effective in helping at least about 75.9% of subjects to achieve 12-month progression-free survival. In some embodiments, the method is effective in helping at least about 89.7% of subjects to achieve 12-month progression-free survival.

[0019] In some embodiments of the various methods or aspects provided herein, the time to complete response or minimal response on the first attempt is in the range of about 0.9 to about 11.1 months. In some embodiments, the time to complete response or minimal response on the first attempt is about 2.1 months on a median basis.

[0020] In some embodiments of the various methods or aspects provided herein, the time to best overall response or best minimal response is approximately 1.1 to approximately 18.6 months. In some embodiments, the median time to best overall response or best minimal response is approximately 6.4 months. In some embodiments, the median time to best overall response or best minimal response is approximately 6.5 months.

[0021] In some embodiments of the various methods or embodiments provided herein, the method further includes treating a subject for adverse events after administering a dose of T cells. In some embodiments, the method includes administering a treatment to a subject to mitigate adverse events. In some embodiments, adverse events include hematological adverse events, non-hematological adverse events, adverse events occurring under the investigational treatment, or any combination thereof. In some embodiments, non-hematological adverse events include infections and / or non-hematological adverse events other than infections. In some embodiments, adverse events include neutropenia, thrombocytopenia, anemia, lymphopenia, upper respiratory tract infection, nasopharyngitis, sinusitis, rhinitis, tonsillitis, pharyngitis, laryngitis, pharyngotonsillitis, COVID-19, COVID-19 pneumonia, asymptomatic COVID-19, neutropenic sepsis, progressive multifocal leukoencephalpathy, septic shock, respiratory failure, pulmonary embolism, lower respiratory tract / lung infection, pneumonia, bronchitis, nausea, hypogammaglobulinemia, diarrhea, fatigue, headache, constipation, hypokalemia, asthenia, peripheral edema, decreased appetite, peripheral sensory neuropathy, back pain, arthralgia, fever, dyspnea, insomnia, or any combination thereof. In some embodiments, adverse events are grade 3 / 4 adverse events. In some embodiments, adverse events persist for more than approximately 30 or 60 days.

[0022] In some embodiments of the various methods or embodiments provided herein, the method further includes treating a subject with respect to a secondary primary malignancy after administering a dose of T cells. In some embodiments, the method includes administering a treatment to a subject to alleviate a secondary primary malignancy. In some embodiments, the secondary primary malignancy includes cutaneous / non-invasive malignancies, hematological malignancies, non-cutaneous / invasive malignancies, or any combination thereof. In some embodiments, the secondary primary malignancy includes basal cell carcinoma, Bowen's disease, squamous cell carcinoma of the lip, malignant melanoma, malignant melanoma in situ, squamous cell carcinoma of the skin, acute myeloid leukemia, myelodysplastic syndrome, peripheral T-cell lymphoma, angiosarcoma, invasive lobular breast carcinoma, pleomorphic malignant fibrous histiocytoma, renal cell carcinoma, tonsil carcinoma, or any combination thereof.

[0023] In some embodiments of the various methods or aspects provided herein, adverse events or secondary primary malignancies occur in a subject at a rate equivalent to that of a subject receiving standard treatment.

[0024] In some embodiments of the various methods or embodiments provided herein, the method further includes treating a subject for CAR-T related adverse events after administering a dose of T cells. In some embodiments, the method includes administering a treatment to a subject to mitigate CAR-T related adverse events. In some embodiments, CAR-T related adverse events include cytokine release syndrome (CRS) and / or neurotoxicity.

[0025] In some embodiments of the various methods or embodiments provided herein, the CAR-T related adverse event is CRS. In some embodiments, CRS occurs in about 60% to about 90% of subjects. In some embodiments, CRS occurs in about 76.1% of subjects. In some embodiments, the maximum toxicity grade of CRS is Grade 1, Grade 2, or Grade 3. In some embodiments, the maximum toxicity grade of CRS is Grade 1. In some embodiments, the maximum toxicity grade of Grade 1 in subjects occurs in about 52.8% of cases. In some embodiments, the maximum toxicity grade of CRS is Grade 2. In some embodiments, the maximum toxicity grade of Grade 2 in subjects occurs in about 22.2% of cases. In some embodiments, the maximum toxicity grade of CRS is Grade 3. In some embodiments, the maximum toxicity grade of Grade 3 in subjects occurs in about 1.1% of cases. In some embodiments, the time to first onset of CRS ranges from about 1 to about 23 days. In some embodiments, the median time to first onset of CRS is about 8 days. In some embodiments, the duration of CRS ranges from about 1 to about 17 days. In some embodiments, the median duration of CRS is about 3 days. In some embodiments of the various methods or embodiments provided herein, treatment of adverse events includes tocilizumab, oxygen, corticosteroids, vasopressors, or any combination thereof.

[0026] In some embodiments of the various methods or aspects provided herein, CAR-T related adverse events are neurotoxic. In some embodiments, neurotoxicity includes immunoeffector cell-associated neurotoxic syndrome or associated symptoms, motor and neurocognitive neurotoxicity, neurotoxic adverse events occurring under study treatment, non-immune effector cell-associated neurotoxic syndrome or associated symptoms, or any combination thereof. In some embodiments, neurotoxicity is immunoeffector cell-associated neurotoxic syndrome or associated symptoms.

[0027] In some embodiments of the various methods or aspects provided herein, immunoeffector cell-associated neurotoxicity syndrome or associated symptoms occur in approximately 4.5% of subjects. In some embodiments, the maximum toxicity grade of immunoeffector cell-associated neurotoxicity syndrome or associated symptoms is Grade 1 or Grade 2. In some embodiments, the maximum toxicity grade of immunoeffector cell-associated neurotoxicity syndrome or associated symptoms is Grade 1. In some embodiments, the maximum toxicity grade of Grade 1 occurs in approximately 3.4% of subjects. In other embodiments, the maximum toxicity grade of immunoeffector cell-associated neurotoxicity syndrome or associated symptoms is Grade 2. In some embodiments, the maximum toxicity grade of Grade 2 occurs in approximately 1.1% of subjects. In some embodiments, the time to onset of immunoeffector cell-associated neurotoxicity syndrome or associated symptoms ranges from approximately 6 to approximately 15 days. In some embodiments, the median time to onset of immunoeffector cell-associated neurotoxicity syndrome or associated symptoms is approximately 9.5 days. In some embodiments, the duration of immunoeffector cell-associated neurotoxicity syndrome or associated symptoms ranges from approximately 1 to approximately 6 days. In some embodiments, the median duration of immunoeffector cell-associated neurotoxicity syndrome or associated symptoms is approximately 2 days. In some embodiments, treatment of adverse events includes corticosteroids and / or tocilizumab.

[0028] In some embodiments of the various methods or aspects provided herein, neurotoxicity is CAR-T cell neurotoxicity. In some embodiments, CAR-T cell neurotoxicity occurs in approximately 17.0% of subjects. In some embodiments, CAR-T cell neurotoxicity includes grade 3 / 4 neurotoxicity, grade 5 neurotoxicity, cranial nerve palsy, peripheral neuropathy, adverse events occurring under study treatment relating to motor and neurocognitive function, or any combination thereof. In some embodiments, CAR-T cell neurotoxicity is grade 3 / 4 neurotoxicity occurring in approximately 2.3% of subjects. In some embodiments, CAR-T cell neurotoxicity is grade 5 neurotoxicity. In some embodiments, CAR-T cell neurotoxicity is cranial nerve palsy. In some embodiments, cranial nerve palsy occurs in approximately 9.1% of subjects. In some embodiments, cranial nerve palsy is grade 2 or grade 3 cranial nerve palsy. In some embodiments, cranial nerve palsy is grade 2 cranial nerve palsy occurring in approximately 8.0% of subjects. In some embodiments, cranial nerve palsy is grade 3, occurring in approximately 1.1% of subjects. In some embodiments, the time from administration of T-cell dose to the onset of cranial nerve palsy ranges from approximately 17 to 60 days. In some embodiments, the median time from administration of T-cell dose to the onset of cranial nerve palsy is approximately 21 days. In some embodiments, cranial nerve palsy occurs in the third, fifth, or seventh cranial nerve. In some embodiments, the duration of cranial nerve palsy ranges from approximately 15 to 262 days. In some embodiments, the median duration of cranial nerve palsy is approximately 77 days. In some embodiments, the treatment includes corticosteroids. In some embodiments, CAR-T cell neurotoxicity is peripheral neuropathy. In some embodiments, peripheral neuropathy occurs in approximately 2.8% of subjects. In some embodiments, peripheral neuropathy is grade 1. In some embodiments, grade 1 peripheral neuropathy occurs in approximately 1.1% of cases. In some embodiments, peripheral neuropathy is grade 2.In some embodiments, grade 2 peripheral neuropathy occurs in approximately 1.1% of cases. In some embodiments, peripheral neuropathy is grade 3. In some embodiments, grade 3 peripheral neuropathy occurs in approximately 0.6% of cases. In some embodiments, CAR-T cell neurotoxicity is an adverse event that occurred during the study treatment for motor and neurocognitive function. In some embodiments, the adverse event that occurred during the study treatment for motor and neurocognitive function was grade 1. In some embodiments, grade 1 adverse events that occurred during the study treatment for motor and neurocognitive function occurred in approximately 0.6% of subjects.

[0029] In some embodiments of the various methods or embodiments provided herein, CD3+ cells containing CAR in the blood of a subject peak about 13 days after administration of T cells to the subject, median. In some embodiments, CD3+ cells containing CAR in the blood of a subject peak at an average concentration of about 1523 cells / μL. In some embodiments, CD3+ cells containing CAR in the blood of a subject remain detectable about 13 to 631 days after administration of T cells to the subject. In some embodiments, CD3+ cells containing CAR in the blood of a subject remain detectable about 57 days after administration of T cells to the subject, median. In some embodiments, the AUC of CD3+ cells containing CAR in the blood of a subject 0-28 The average value is approximately 12,504 cells / μL.

[0030] In some embodiments of the various methods or embodiments provided herein, the first VHH domain includes CDR1, CDR2, and CDR3 of VHH domains containing the amino acid sequence of SEQ ID NO: 2, and the second VHH domain includes CDR1, CDR2, and CDR3 of VHH domains containing the amino acid sequence of SEQ ID NO: 4. In some embodiments, the first VHH domain includes CDR1 containing the amino acid sequence of SEQ ID NO: 18, CDR2 containing the amino acid sequence of SEQ ID NO: 19, and CDR3 containing the amino acid sequence of SEQ ID NO: 20; and the second VHH domain includes CDR1 containing the amino acid sequence of SEQ ID NO: 21, CDR2 containing the amino acid sequence of SEQ ID NO: 22, and CDR3 containing the amino acid sequence of SEQ ID NO: 23. In some embodiments, the first VHH domain includes the amino acid sequence of SEQ ID NO: 2, and the second VHH domain includes the amino acid sequence of SEQ ID NO: 4. In some embodiments, the first VHH domain is at the N-terminus of the second VHH domain. In some embodiments, the first VHH domain is located at the C-terminus of the second VHH domain. In some embodiments, the first VHH domain is linked to the second VHH domain via a linker containing the amino acid sequence of SEQ ID NO: 3. In some embodiments, the transmembrane domain is derived from a molecule selected from the group consisting of CD8α, CD4, CD28, CD137, CD80, CD86, CD152, and PD1. In some embodiments, the transmembrane domain is derived from CD8α and contains the amino acid sequence of SEQ ID NO: 6. In some embodiments, the intracellular signaling domain includes the primary intracellular signaling domain of an immunoeffector cell. In some embodiments, the primary intracellular signaling domain is derived from CD3ζ containing the amino acid sequence of SEQ ID NO: 8. In some embodiments, the intracellular signaling domain includes a co-stimulatory signaling domain. In some embodiments, the co-stimulatory signaling domain is derived from a co-stimulatory molecule selected from the group consisting of ligands CD27, CD28, CD137, OX40, CD30, CD40, CD3, LFA-1, ICOS, CD2, CD7, LIGHT, NKG2C, B7-H3, CD83, and any combination thereof. In some embodiments, the co-stimulatory signaling domain includes the cytoplasmic domain of CD137 containing the amino acid sequence of SEQ ID NO: 7.In some embodiments, the CAR further includes a hinge domain located between the C-terminus of the extracellular antigen-binding domain and the N-terminus of the transmembrane domain. In some embodiments, the hinge domain is derived from CD8α containing the amino acid sequence of SEQ ID NO: 5. In some embodiments, the CAR further includes a signal peptide located at the N-terminus of the polypeptide. In some embodiments, the signal peptide is derived from CD8α containing the amino acid sequence of SEQ ID NO: 1. In some embodiments, the CAR contains the amino acid sequence of SEQ ID NO: 17.

[0031] In some embodiments, the Specified Description provides a method for treating a subject having multiple myeloma, comprising administering to the subject a certain dose of chimeric antigen receptor (CAR)-containing T cells comprising (a) an extracellular antigen-binding domain having the ability to specifically bind to an epitope of B cell maturation antigen (BCMA), (b) a transmembrane domain, and (c) an intracellular signaling domain, wherein the subject has received one to three prior lines of treatment, including immunomodulatory drug (IMiD) therapy, and is resistant to IMiD. In some embodiments, administration of the dose of T cells reduces the subject's risk of disease progression or death. In some embodiments, the risk of disease progression or death is reduced compared to administration of daratumumab-pomalidomide-dexamethasone (DPd) or pomalidomide-bortezomib-dexamethasone (PVd) treatment. In some embodiments, the risk of disease progression or death is reduced compared to standard of care (SOC) therapy, which includes either daratumumab-pomalidomide-dexamethasone (DPd) therapy or pomalidomide-bortezomib-dexamethasone (PVd) therapy. In some embodiments, the risk of disease progression or death is reduced compared to ide-cel therapy. In some embodiments, subjects exhibit a disease progression or death risk reduced by approximately 60% to approximately 75%. In some embodiments, subjects exhibit a disease progression or death risk reduced by approximately 74%.

[0032] In some embodiments, the Specified Description provides a method for treating a subject having multiple myeloma, comprising administering to the subject a certain dose of chimeric antigen receptor (CAR)-containing T cells comprising (a) an extracellular antigen-binding domain having the ability to specifically bind to a B-cell maturation antigen (BCMA) epitope, (b) a transmembrane domain, and (c) an intracellular signaling domain, wherein the subject has received one to three prior treatment lines, including immunomodulatory drug (IMiD) therapy, is resistant to IMiD, and the administration of the dose of T cells is more effective in achieving a best partial response (VGPR) or better in the subject compared to administration of DPd or PVd therapy. In some embodiments, the VGPR or better rate after administration of the treatment is approximately 81.3%. In some embodiments, the VGPR or better rate after administration of DPd or PVd is approximately 45.5%. In some embodiments, the VGPR or better rate after administration of standard of care (SOC) therapy, including administration of either DPd or PVd, is approximately 45.5%. In some embodiments, the treatment is more effective in achieving a strict complete response (sCR) in subjects compared to administration of DPd or PVd therapy. In some embodiments, the sCR rate after administration of the treatment is approximately 58.2%. In some embodiments, the sCR rate after administration of DPd or PVd is approximately 15.2%.

[0033] Another aspect of the present disclosure is a method for treating a subject having multiple myeloma, comprising administering to the subject a certain dose of chimeric antigen receptor (CAR)-containing T cells comprising (a) an extracellular antigen-binding domain having the ability to specifically bind to an epitope of B cell maturation antigen (BCMA), (b) a transmembrane domain, and (c) an intracellular signaling domain, wherein the subject has received one to three prior lines of treatment, including immunomodulatory drug (IMiD) therapy, and is resistant to IMiD.

[0034] In some embodiments, the subjects exhibit a reduced risk of CAR-T related adverse events, which optionally include cytokine release syndrome (CRS) and / or neurotoxicity.

[0035] In some embodiments, the CAR-T-related adverse event is CRS, and optionally, CRS occurs in 60% to approximately 90% or approximately 76.1% of subjects, and optionally, the maximum toxicity grade of CRS is Grade 1, Grade 2, or Grade 3.

[0036] In some embodiments, the CAR-T-related adverse event is CRS, and optionally, CRS occurs in approximately 60% to 90% or 76.1% of subjects, and optionally, the maximum toxicity grade of CRS is Grade 1, Grade 2, or Grade 3, and optionally, the maximum toxicity grade of CRS is Grade 1, and optionally, the maximum toxicity grade of Grade 1 occurs in approximately 52.8% of subjects.

[0037] In some embodiments, the CAR-T-related adverse event is CRS, and optionally, CRS occurs in approximately 60% to 90% or 76.1% of subjects, and optionally, the maximum toxicity grade of CRS is Grade 1, Grade 2, or Grade 3, and optionally, the maximum toxicity grade of CRS is Grade 2, and optionally, Grade 2 occurs in approximately 22.2% of subjects.

[0038] In some embodiments, the CAR-T-related adverse event is CRS, and optionally, CRS occurs in approximately 60% to 90% or 76.1% of subjects, and optionally, the maximum toxicity grade of CRS is Grade 1, Grade 2, or Grade 3, and optionally, the maximum toxicity grade of CRS is Grade 3, and optionally, the maximum toxicity grade of Grade 3 occurs in approximately 1.1% of subjects.

[0039] In some embodiments, the CAR-T-related adverse event is CRS, and optionally, CRS occurs in approximately 60% to 90% or 76.1% of subjects, optionally, the maximum toxicity grade of CRS is Grade 1, Grade 2, or Grade 3, optionally, the time to the first onset of CRS is in the range of approximately 1 to 23 days, optionally, the median time to the first onset of CRS is approximately 8 days.

[0040] In one embodiment, the CAR-T-related adverse event is CRS, and optionally, CRS occurs in approximately 60% to 90% or 76.1% of subjects, and optionally, the maximum toxicity grade of CRS is Grade 1, Grade 2, or Grade 3, and optionally, the duration of CRS is in the range of approximately 1 to 17 days, and optionally, the median duration of CRS is approximately 3 days.

[0041] In some embodiments, the CAR-T related adverse event is CRS, and optionally, CRS occurs in approximately 60% to approximately 90% or approximately 76.1% of subjects, and optionally, the maximum toxicity grade of CRS is grade 1, grade 2 or grade 3, and optionally, the method further comprises administering tocilizumab, oxygen, corticosteroids, vasopressors, or any combination thereof to the subjects.

[0042] In some embodiments, CAR-T related adverse events are neurotoxic, and optionally, neurotoxicity includes immunoeffector cell-associated neurotoxic syndrome or associated symptoms, motor / neurocognitive neurotoxicity, neurotoxic adverse events occurring under study treatment, non-immune effector cell-associated neurotoxic syndrome or associated symptoms, or any combination thereof, and optionally, neurotoxicity is immunoeffector cell-associated neurotoxic syndrome or associated symptoms.

[0043] In some embodiments, CAR-T related adverse events are neurotoxic, and optionally, neurotoxicity includes immunoeffector cell-associated neurotoxic syndrome or associated symptoms, motor / neurocognitive neurotoxicity, neurotoxic adverse events occurring under study treatment, non-immune effector cell-associated neurotoxic syndrome or associated symptoms, or any combination thereof, and optionally, neurotoxicity is immunoeffector cell-associated neurotoxic syndrome or associated symptoms, and optionally, immunoeffector cell-associated neurotoxic syndrome or associated symptoms occur in approximately 4.5% of subjects.

[0044] In some embodiments, CAR-T related adverse events are neurotoxic, and optionally, the neurotoxicity includes immunoeffector cell-associated neurotoxic syndrome or associated symptoms, motor / neurocognitive neurotoxicity, neurotoxic adverse events occurring under study treatment, non-immune effector cell-associated neurotoxic syndrome or associated symptoms, or any combination thereof; optionally, the neurotoxicity is immunoeffector cell-associated neurotoxic syndrome or associated symptoms, and optionally, the maximum toxicity grade of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is Grade 1 or Grade 2; optionally, the maximum toxicity grade of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is Grade 1, and optionally, the maximum toxicity grade of Grade 1 in subjects occurs at a rate of approximately 3.4%; or optionally, the maximum toxicity grade of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is Grade 2, and optionally, the maximum toxicity grade of Grade 2 in subjects occurs at a rate of approximately 1.1%.

[0045] In some embodiments, CAR-T related adverse events are neurotoxic, and optionally, neurotoxicity includes immunoeffector cell-associated neurotoxic syndrome or associated symptoms, motor / neurocognitive neurotoxicity, neurotoxic adverse events occurring under study treatment, non-immune effector cell-associated neurotoxic syndrome or associated symptoms, or any combination thereof; optionally, neurotoxicity is immunoeffector cell-associated neurotoxic syndrome or associated symptoms; further optionally, the time to onset of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is in the range of approximately 6 to approximately 15 days; and optionally, the median time to onset of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is approximately 9.5 days.

[0046] In some embodiments, CAR-T related adverse events are neurotoxic, and optionally, neurotoxicity includes immunoeffector cell-associated neurotoxic syndrome or associated symptoms, motor / neurocognitive neurotoxicity, neurotoxic adverse events occurring under study treatment, non-immune effector cell-associated neurotoxic syndrome or associated symptoms, or any combination thereof; optionally, neurotoxicity is immunoeffector cell-associated neurotoxic syndrome or associated symptoms; further optionally, the duration of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is in the range of approximately 1 to approximately 6 days; and optionally, the median duration of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is approximately 2 days.

[0047] In some embodiments, CAR-T related adverse events are neurotoxic, and optionally, neurotoxicity includes immunoeffector cell-associated neurotoxic syndrome or associated symptoms, motor / neurocognitive neurotoxicity, neurotoxic adverse events occurring under study treatment, non-immune effector cell-associated neurotoxic syndrome or associated symptoms, or any combination thereof; optionally, neurotoxicity is immunoeffector cell-associated neurotoxic syndrome or associated symptoms; and optionally, treatment includes corticosteroids and / or tocilizumab.

[0048] Another aspect of the present disclosure is a method for treating a subject having multiple myeloma, comprising administering to the subject a certain dose of chimeric antigen receptor (CAR)-containing T cells comprising (a) an extracellular antigen-binding domain having the ability to specifically bind to an epitope of B cell maturation antigen (BCMA), (b) a transmembrane domain, and (c) an intracellular signaling domain, wherein the subject has received one to three prior lines of treatment, including immunomodulatory drug (IMiD) therapy, and is resistant to IMiD.

[0049] In some embodiments, subjects exhibit a reduced risk of CAR-T related adverse events, and optionally, CAR-T related adverse events include CAR-T cell neurotoxicity, and optionally, CAR-T cell neurotoxicity occurs in approximately 17.0% of subjects, and optionally, CAR-T cell neurotoxicity includes grade 3 / 4 neurotoxicity, grade 5 neurotoxicity, cranial nerve palsy, peripheral neuropathy, adverse events related to motor / neurocognitive function that occurred under study treatment, or any combination thereof.

[0050] In some embodiments, CAR-T cell neurotoxicity is a grade 3 / 4 neurotoxicity occurring in approximately 2.3% of subjects.

[0051] In some embodiments, CAR-T cell neurotoxicity is grade 5 neurotoxicity.

[0052] In some embodiments, CAR-T cell neurotoxicity is cranial nerve palsy, which occurs in approximately 9.1% of subjects, with optional selection.

[0053] In some embodiments, CAR-T cell neurotoxicity is cranial palsy, which occurs in approximately 9.1% of subjects at random selection, and is of grade 2 or grade 3, and further at random selection, is grade 2 cranial palsy occurring in approximately 8.0% of subjects, and further at random selection, is grade 3 cranial palsy occurring in approximately 1.1% of subjects.

[0054] In some embodiments, CAR-T cell neurotoxicity is cranial nerve palsy, which occurs in approximately 9.1% of subjects in an optional setting. The time from administration of T cell doses to the onset of cranial nerve palsy ranges from approximately 17 to 60 days, and in an optional setting, the median time from administration of T cell doses to the onset of cranial nerve palsy is approximately 21 days.

[0055] In some embodiments, CAR-T cell neurotoxicity is cranial nerve palsy, which occurs in approximately 9.1% of subjects at an optional rate, and the cranial nerve palsy develops in the third, fifth, or seventh cranial nerve.

[0056] In some embodiments, CAR-T cell neurotoxicity is cranial nerve palsy, which occurs in approximately 9.1% of subjects in an optional selection, with a duration of approximately 15 to 262 days, and further optional selection, with a median duration of approximately 77 days.

[0057] In some embodiments, CAR-T cell neurotoxicity is cranial nerve palsy, which occurs in approximately 9.1% of subjects at an optional rate, and the method further comprises administering a treatment containing a corticosteroid to the subjects.

[0058] In some embodiments, CAR-T cell neurotoxicity is peripheral neuropathy, and in optional selection, peripheral neuropathy occurs in approximately 2.8% of subjects. In some embodiments, peripheral neuropathy is Grade 1. In some embodiments, Grade 1 peripheral neuropathy occurs in approximately 1.1% of subjects. In some embodiments, peripheral neuropathy is Grade 2. In some embodiments, Grade 2 peripheral neuropathy occurs in approximately 1.1% of subjects. In some embodiments, peripheral neuropathy is Grade 3. In some embodiments, Grade 3 peripheral neuropathy occurs in approximately 0.6% of subjects.

[0059] In some embodiments, CAR-T cell neurotoxicity is an adverse event that occurs during the study treatment for motor and neurocognitive function. In random selection, the adverse event that occurs during the study treatment for motor and neurocognitive function is grade 1, and in random selection, grade 1 adverse events that occur during the study treatment for motor and neurocognitive function occur in approximately 0.6% of subjects.

[0060] Features described in the context of different aspects and embodiments of this disclosure may be used together and / or interchangeable with one another. Similarly, features described in the context of a single embodiment may also be provided separately or in any preferred partial combination. However, all methods described herein, however expressed, may be described as corresponding uses, in particular as medical uses. [Brief explanation of the drawing]

[0061] [Figure 1] This shows the expression of BCMA antigens on the surface of GC, memory and plasmablasts in lymph nodes, long-lived plasma cells in bone marrow LN and MALT, and on multiple myeloma cells. BAFF-R antigen is not expressed on plasmablasts, long-lived plasma cells, or multiple myeloma cells. TACI is expressed on memory and plasmablasts, long-lived plasma cells, and multiple myeloma cells. CD138 is expressed only on long-lived plasma cells and multiple myeloma cells.

[0062] [Figure 2] This document describes the design of siltacaptagene autoleucel, a CAR. Siltacaptagene autoleucel contains two VHH domains, in contrast to the single VL domain and single VH domain found on various other CARs. Siltacaptagene autoleucel also contains intracellular CD137 and human CD3 zeta domains.

[0063] [Figure 3]A schematic diagram is shown illustrating the preparation of a virus encoding siltacaptagene autoleucel CAR, the transduction of the virus into patient-derived T cells, and the subsequent preparation of CAR T cells expressing siltacaptagene autoleucel.

[0064] [Figure 4] A schematic diagram of the study design for siltacabutagen autoleucel CAR T cells is shown. The patient population includes patients with relapsed or resistant multiple myeloma who have received 1 to 3 prior treatment lines, including immunomodulatory agents, or who are biresistant to PI / IMiD and have a history of exposure to PI, IMiD, and anti-CD38. The primary objective is to compare the efficacy and safety of siltacabutagen autoleucel CAR T cells in the aforementioned patient population in a randomized controlled trial (Phase 3) with physician's choice between two highly effective standard therapies.

[0065] [Figure 5] This graph shows the allocation of research subjects in each treatment group.

[0066] [Figure 6A-6C] The Kaplan-Meier intention-based analysis is shown. Figure 6A shows progression-free survival by treatment group. Figure 6B shows progression-free survival by treatment group, stratified by the number of prior treatment lines. Figure 6C shows overall survival by treatment group.

[0067] [Figure 7]Forest plots of subgroup analyses for progression-free survival are shown. Abbreviations: cilta-cel, siltacabutagene autoleucel; DPd, daratumumab-pomalidomide-dexamethasone; ECOG, Eastern Cooperative Oncology Group; IMID, immunomodulatory drugs; ISS, International Staging Classification; MM, multiple myeloma; NCI, National Cancer Institute; PI, proteasome inhibitors; PVd, pomalidomide-bortezomib-dexamethasone; SOC, standard care. a Hazard ratios and 95% CIs from a Cox proportional hazards model with treatment as the sole explanatory variable, including only PFS events occurring more than 8 weeks after randomization. Hazard ratios less than 1 indicate a favorable cilta-cel arm. b Based on web automated response system randomization strata. c Based on serum β-2 microglobulin and albumin. d Based on subjects with measurable disease in serum. eFISH testing is positive for del(17p), t(14;16), t(4;14), and / or gain / amp(1q). High-risk cytogenetics, as defined by the protocol, refers to any of the four marker abnormalities. Based on the Modified Dietary Rehabilitation (MDRD) formula for renal disease.

[0068] [Figures 8A-8D] Figure 8A shows the PFS and OS response rates of patients who received cilta-cel in CARTITUDE-1 and CARTITUDE-4. Figure 8B shows the PFS of patients who received cilta-cel as a research treatment in CARTITUDE-1. Figure 8C shows the OS of patients who received cilta-cel as a research treatment in CARTITUDE-1. Figure 8D shows the OS of patients who received cilta-cel as a research treatment in CARTITUDE-4. [Modes for carrying out the invention]

[0069] This disclosure also provides related nucleic acids, recombinant expression vectors, host cells, cell populations, antibodies or their antigen-binding moieties, and pharmaceutical compositions relating to immune cells and CAR-expressing T cells. Dosage regimens and forms of administration, as well as methods of treatment with CAR-T cells, are also provided.

[0070] Some aspects and embodiments of this disclosure are described below with respect to examples for illustrative purposes only. It should be understood that many specific details, relevances, and methods are described to provide a full understanding of this disclosure. However, those skilled in the art will readily recognize that this disclosure may be practiced without one or more specific details, or using other methods, protocols, reagents, cell lines, and animals. Since some actions may be performed in a different order and / or concurrently with other actions or events, this disclosure is not limited by the illustrated order of actions or events. Furthermore, not all illustrated actions, processes, or events are required to implement the methodologies described herein.

[0071] Unless otherwise defined, all technical terms, notations, and other scientific or academic terms used herein are intended to have meanings that are generally understood by those skilled in the art to which this disclosure relates. In some cases, terms that have generally understood meanings are defined herein for clarity and / or for easy reference, and the inclusion of such definitions herein should not necessarily be interpreted as representing a substantial difference from those generally understood in the art. Furthermore, terms such as those defined in commonly used dictionaries should be interpreted as having meanings consistent with their meanings in the context of the relevant art and / or as they are otherwise defined herein.

[0072] The terms "about" or "approximately" include being within a statistically meaningful range from a given value. Such a range may be within a certain order of magnitude from a given value or range, preferably within 50%, more preferably within 20%, even more preferably within 10%, and still more preferably within 5%. The acceptable variation included in the terms "about" or "approximately" depends on the detailed system under study and will be readily apparent to those skilled in the art.

[0073] The term "antibody" includes monoclonal antibodies (including full-length quadruple-chain antibodies or full-length heavy-chain-only antibodies with an immunoglobulin Fc region), antibody compositions exhibiting multi-epitope specificity, multispecific antibodies (e.g., bispecific antibodies, diabodies, and single-chain molecules), and antibody fragments (e.g., Fab, F(ab')2, and Fv). The term "immunoglobulin" (Ig) is used herein as synonymous with "antibody." Antibodies as intended herein include single-domain antibodies, such as heavy-chain-only antibodies.

[0074] The term "heavy-chain-only antibody" or "HCAb" refers to a functional antibody that contains a heavy chain but lacks the light chain typically found in quadruple-chain antibodies. Camelids (such as camels, llamas, or alpacas) are known to produce HCAbs.

[0075] The term "single-domain antibody" or "sdAb" refers to a single antigen-binding polypeptide having three complementarity-determining regions (CDRs). An sdAb possesses the ability to bind to an antigen independently, without being paired with a corresponding CDR-containing polypeptide. In some cases, single-domain antibodies are engineered from camelid HCAbs, whose heavy-chain variable domains are referred to as "VHHs." Some VHHs may also be known as "nanobodies." Camelid sdAbs are among the smallest known antigen-binding antibody fragments (see, e.g., Hamers-Casterman et al., Nature 363:446-8 (1993); Greenberg et al., Nature 374:168-73 (1995); Hassanzadeh-Ghassabeh et al., Nanomedicine (Lond), 8:1013-26 (2013)). The basic VHH has the following structure from the N-terminus to the C-terminus: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4, where FR1 to FR4 refer to framework regions 1 to 4, respectively, and CDR1 to CDR3 refer to complementarity determination regions 1 to 3.

[0076] The "variable region" or "variable domain" of an antibody refers to the amino-terminal domain of the heavy or light chain of the antibody. The variable domains of the heavy and light chains may be referred to as "VH" and "VL," respectively. These domains are generally the most variable parts of the antibody (compared to other antibodies of the same class) and contain the antigen-binding site. Only heavy chain antibodies from camelid species have a single heavy chain variable region, which is referred to as the "VHH" domain. Therefore, VHH is a special type of variable region.

[0077] The term "variable" refers to the fact that specific segments of the variable domain differ extensively in sequence between antibodies. The V domain (i.e., the variable domain) mediates antigen binding and defines the specificity of a particular antibody to that particular antigen. However, its variability is not evenly distributed across the entire width of the variable domain. Instead, variability is concentrated in three segments called hypervariable regions (HVRs) in both the light chain and heavy chain variable domains. The more highly conserved portions within the variable domain are called framework regions (FRs). The variable domains of the natural heavy and light chains each contain four FR regions that generally take on a β-sheet configuration, and the three HVRs linking them form loops that link, or sometimes form part of, the β-sheet structure. The HVRs on each chain are held together in close proximity by the FR region, contributing to the formation of the antigen-binding site of the antibody (or, if the antibody is not an sdAb or HCAb, with the HVR from the other chain) (see Kabat et al., Sequences of Immunological Interest, Fifth Edition, National Institute of Health, Bethesda, Md. (1991)). The constant domain does not directly participate in antibody binding to the antigen, but exhibits various effector functions, such as the antibody's involvement in antibody-dependent cell-mediated cytotoxicity.

[0078] The terms “antibody fragment,” “antibody fragment,” “functional antibody fragment,” and “antigen-binding moiety” are used herein synonymously to mean one or more fragments or portions of an antibody that possess the ability to specifically bind to an antigen (see Holliger et al., Nat. Biotech., 23(9):1 126-1129 (2005) for general purposes). The antigen-recognizing moiety of a CAR encoded by a nucleic acid sequence disclosed herein may include any BCMA-conjugated antibody fragment. The antibody fragment preferably includes, for example, one or more CDRs, variable regions (or portions thereof), constant regions (or portions thereof), or a combination thereof. Examples of antibody fragments include, but are not limited to, (i) Fab fragments, which are monovalent fragments consisting of VL, VH, CL, and CHI domains; (ii) F(ab')2 fragments, which are bivalent fragments containing two Fab fragments linked by disulfide crosslinks at the hinge region; (iii) Fv fragments, which consist of the VL and VH domains of a single arm of the antibody; and (iv) single-chain Fv(scFv) monovalent molecules, which consist of two domains (i.e., VL and VH) of an Fv fragment linked by a synthetic linker that allows them to be synthesized as a single polypeptide chain (e.g., Bird et al., Science, 242:423-426 (1988); Huston et al., Proc. Natl. Acad. Sci. USA, 85:5879-5883 (1988); and Osbourn et al. (See al., Nat. Biotechnol, 16:778 (1998)) and (v) dimers of polypeptide chains, wherein each polypeptide chain contains VH linked to VL by a peptide linker, and the linker is too short for pairing between VH and VL on the same polypeptide chain, thereby promoting pairing between complementary domains on different VH-VL polypeptide chains to produce a dimer molecule having two functional antigen-binding sites. Antibody fragments are known in the art and are described in detail, for example, in U.S. Patent Application Publication No. 2009 / 0093024 A1.

[0079] As used herein, the terms “specifically bind,” “specifically recognize,” or “specific to” refer to measurable and reproducible interactions that can determine the presence of a target in the presence of a heterogeneous population of molecules, including biomolecules, such as binding between a target and an antigen-binding protein (such as a CAR or VHH).

[0080] The term "specificity" refers to the selective recognition of an antigen-binding protein (such as CAR or VHH) for a specific epitope of an antigen. Natural antibodies, for example, are monospecific.

[0081] A "chimeric antigen receptor" or "CAR" is an artificially constructed hybrid protein or polypeptide having an antigen-binding domain of an antibody (or antibody fragment) linked to a T cell signaling domain. A key characteristic of CARs is their ability to redirect T cell specificity and reactivity to a non-MHC-restricted target of choice by utilizing the antigen-binding properties of a monoclonal antibody. This non-MHC-restricted antigen recognition allows CAR-expressing T cells to recognize antigens independently of antigen processing, thus circumventing major tumor evasion mechanisms. Furthermore, when expressed on T cells, advantageously, CARs do not dimerize with the endogenous T cell receptor (TCR) α and β chains. T cells expressing CARs are referred to herein as CAR T cells, CAR-T cells, or CAR-modified T cells, and these terms are used synonymously herein. These cells can be genetically modified to stably express an antibody-binding domain on their surface, conferring novel MHC-independent antigen specificity. "BCMA CAR" refers to a CAR having an extracellular binding domain specific to BCMA. A "biepitope CAR" refers to a CAR that has extracellular binding domains specific to two different epitopes on the BCMA.

[0082] Silta-cabutagen autoleucel ("cilta-cel") is a chimeric antigen receptor T cell (CAR-T) therapy containing two B cell maturation antigen (BCMA)-targeted VHH domains designed to confer avidity to BCMA. Cilta-cel may contain T lymphocytes transduced with the silta-cabutagen autoleucel CAR, which is encoded by a lentiviral vector. This CAR targets human B cell maturation antigen (BCMA CAR). A diagram of the lentiviral vector encoding the cilta-cel CAR is provided in Figure 2. The amino acid sequence of the cilta-cel CAR is the amino acid sequence of Sequence ID No. 17.

[0083] The terms "expression" and "expression" mean enabling or causing a state in which information in a gene or DNA sequence is produced. For example, expression can take the form of producing a protein by activating cellular functions involved in the transcription and translation of the corresponding gene or DNA sequence. When a DNA sequence is expressed in or by a cell, "expression products," such as proteins, are formed. Expression products themselves, such as the resulting proteins, are also said to be "expressed" by the cell. Expression products can be characterized as intracellular, extracellular, or transmembrane.

[0084] The term “to treat” or “treatment” refers to a therapeutic action aimed at delaying or reducing an undesirable physiological change or disease, or providing a beneficial or desired clinical outcome during treatment. Beneficial or desired clinical outcomes include, whether detectable or undetectable, relief of symptoms, reduction of disease severity, stabilization of the disease state (i.e., no worsening), delay or slowing of disease progression, improvement or mitigation of the disease state, and / or remission (whether partial or complete). “Treatment” may also mean extending survival compared to the survival expected if the subject did not receive treatment. Those in need of treatment include subjects who already have an undesirable physiological change or disease, as well as subjects who are susceptible to physiological changes or disease. Treatment may involve therapeutic agents, also referred to herein as “drugs” or “pharmacotherapy,” which may be intended to help achieve a desired or beneficial clinical outcome through their action. Therapeutic agents or drugs may be administered to a subject by numerous routes, including at least intravenous and oral routes. The term "intravenous" refers, in relation to the administration of a therapeutic agent or drug, to the administration of the said therapeutic agent or drug through one or more veins. The term "oral" refers, in relation to the administration of a therapeutic agent or drug, to the administration of the said therapeutic agent or drug through a pathway such as the mouth or oral cavity.

[0085] As used herein, the term “subject” refers to an animal. The terms “subject” and “patient” may be used synonymously herein to refer to a subject. Therefore, “subject” includes a disease or a human being under treatment as a patient for the prevention of a disease. The methods described herein may be used to treat animal subjects belonging to any classification. Examples of such animals include mammals. Examples of mammals include, but are not limited to, rodents such as mice and hamsters, and mammals of the order Lagomorpha such as rabbits. Mammals may also be of the order Carnivora, including felines (cats) and canines (dogs). Mammals may also be of the order Artiodactyla, including bovines (cats) and pigs, or of the order Persodactyla, including equids (horses). Mammals may be of the order Primates, Ceboids, or Simoids (monkeys), or of the order Anthropoids (humans and apes). In some embodiments, the mammal is a human.

[0086] The term "effective" as applied to dosage or quantity refers to the amount of compound or pharmaceutical composition sufficient to produce the desired activity when administered to the subject in need. Note that when administering a combination of active ingredients, the effective dose of that combination may or may not include the amounts of each ingredient that would have been effective if administered individually. The exact amount required will vary from subject to subject, depending on the species, age, and overall condition of the subject, the severity of the condition being treated, the specific one or more medications used, and the mode of administration.

[0087] When used in connection with the compositions described herein, the term “pharmaceutically acceptable” refers to molecular entities and other components of such compositions that are physiologically tolerable upon administration to mammals (e.g., humans) and that typically do not produce adverse reactions. Preferably, the term “pharmaceutically acceptable” means that the use in mammals, and more specifically in humans, is approved by federal or state regulatory authorities, or is listed in the United States Pharmacopeia or other generally accepted pharmacopoeias.

[0088] The term “treatment line,” when used in connection with the treatment methods herein, refers to one or more planned cycles of monotherapy or combination therapy, and one or more cycles of a planned treatment program, which may consist of a series of treatments administered in a planned manner. For example, a planned treatment approach of autologous stem cell transplantation followed by induction therapy and subsequent maintenance constitutes one treatment line. A new treatment line is considered initiated when a planned course of treatment is modified to include other therapeutic agents or medications (alone or in combination) as a result of disease progression, relapse, or toxicity. A new treatment line is also considered initiated when a planned period of observation-free treatment is interrupted by the need for additional treatment for the disease.

[0089] The term "resistant," when used in connection with treatment with a particular therapeutic agent or pharmacopoeia as herein, refers to a disease or disease subject that is unable to respond to such therapeutic agent or pharmacopoeia. The phrase "resistant myeloma" refers to multiple myeloma that is unresponsive during first-line therapy or salvage therapy, or that has progressed within 60 days of the last therapy.

[0090] The term "non-responsive disease" refers to either the failure to achieve minimal response or the development of a progressive disease during therapy.

[0091] The term "hazard ratio" refers to a measure of the relative rate of progression to the endpoint compared to a control group. In outcome-based clinical trials, a reduction in the hazard ratio of a trial arm compared to the control indicates that the treatment used in the trial arm reduces the risk of the endpoint, in the cases described herein, the risk of disease progression or death. Preferably, the hazard ratio is calculated by a stratified constant piecewise weighted log-rank test.

[0092] The terminology used herein is intended solely to describe specific aspects or embodiments and is not intended to limit them. When used herein, the indefinite articles "a," "an," and "the" should be understood to include multiple references unless specifically indicated by the context.

[0093] Throughout this disclosure, various aspects and embodiments of the disclosure may be presented in the form of scopes. It should be understood that the use of scopes is solely for convenience and brevity and should not be interpreted as a definitive limitation of the scope of the disclosure. Accordingly, scope descriptions should be interpreted as specifically disclosing all possible sub-scopes and the individual numerical values ​​within those scopes. For example, a scope description such as 1-6 should be considered to specifically disclose sub-scopes such as 1-3, 1-4, 1-5, 2-4, 2-6, 3-6, and the individual numerical values ​​within those scopes, such as 1, 2, 2.7, 3, 4, 5, 5.3, and 6. As another example, a scope such as 95-99% identity includes those having 95%, 96%, 97%, 98%, or 99% identity, and includes sub-scopes such as 96-99%, 96-98%, 96-97%, 97-99%, 97-98%, and 98-99% identity. This applies regardless of the range.

[0094] vector The polynucleotide sequences encoding CARs described herein can be obtained using standard recombination techniques. The desired polynucleotide sequences can be isolated from antibody-producing cells, such as hybridoma cells, and sequenced. Alternatively, polynucleotides can be synthesized using a nucleotide synthesizer or PCR techniques.

[0095] This disclosure also provides vectors comprising nucleic acid sequences encoding the CARs disclosed herein. The vectors may be, for example, plasmids, cosmids, viral vectors (e.g., retroviral or adenoviral), or phages. Suitable vectors and methods for vector preparation are well known in the art (see, for example, Sambrook et al. and Ausubel et al.).

[0096] In addition to the nucleic acid sequences encoding the CARs disclosed herein, the vectors preferably include expression regulatory sequences such as promoters, enhancers, polyadenylation signals, transcription terminators, and intra-sequence ribosome entry sites (IRESs) that result in the expression of the nucleic acid sequences in host cells. Exemplary expression regulatory sequences are known in the art and are described, for example, in Goeddel, Gene Expression Technology: Methods in Enzymology, Vol. 185, Academic Press, San Diego, Calif. (1990).

[0097] In some embodiments, the vector includes a promoter. A large number of promoters recognized by various potential host cells are well known. A selected promoter can be operably ligated to the cistron DNA encoding the CAR disclosed herein by extracting the promoter from source DNA by restriction enzyme digestion and inserting the isolated promoter sequence into the vector of this application. A large number of promoters, including constitutive, inducible, and repressible promoters from various different sources, are well known in the art. Typical sources of promoters include, for example, viruses, mammals, insects, plants, yeasts, and bacteria, and suitable promoters from these sources are readily available or can be synthetically produced based on sequences publicly available from repositories such as ATCC and other commercial or individual sources. Promoters can be unidirectional (i.e., initiating transcription in one direction) or bidirectional (i.e., initiating transcription in either the 3' or 5' direction). Non-limiting examples of promoters include, for example, the T7 bacterial expression system, the pBAD(araA) bacterial expression system, the cytomegalovirus (CMV) promoter, the SV40 promoter, and the RSV promoter. Examples of inducible promoters include the Tet system (US Patent Nos. 5,464,758 and 5,814,618), the ecdysone-inducible system (No et al., Proc. Natl. Acad. Sci., 93:3346-3351 (1996)), the T-REX® system (Invitrogen, Carlsbad, CA), the LACSWITCH® system (Stratagene, San Diego, CA), and the Cre-ERT tamoxifen-inducible recombinase system (Indra et al., Nuc. Acid. Res., 27:4324-4327 (1999); Nuc. Acid. Res., 28:e99 (2000); US Patent No. 7,112,715; and Kramer & Fussenegger, Methods). Mol. Biol, 308:123-144 (2005) is one example.

[0098] In some embodiments, the vector includes an “enhancer.” The term “enhancer,” as used herein, refers, for example, to a DNA sequence that increases the transcription of a operably linked nucleic acid sequence. Enhancers may be located several kilobases away from the coding region of the nucleic acid sequence and may mediate regulatory binding, DNA methylation patterns, or changes in DNA structure. Numerous enhancers from various different sources are well known in the Art and are available as or within cloned polynucleotides (e.g., from repositories such as ATCC and other commercial or personal sources). Some polynucleotides containing promoters (such as the commonly used CMV promoter) also include enhancer sequences. Enhancers may be located upstream, within, or downstream of the coding sequence. The term “Ig enhancer” refers to an enhancer element derived from an enhancer region mapped within an immunoglobulin (Ig) locus. Examples of such Ig enhancers include heavy-chain (mu) 5' enhancers, light-chain (kappa) 5' enhancers, kappa and mu intron enhancers, and 3' enhancers (see, for example, Paul WE (ed), Fundamental Immunology, 3rd Edition, Raven Press, New York (1993), pages 353-363; and U.S. Patent No. 5,885,827).

[0099] In some embodiments, the vector includes a “selection marker gene.” The term “selection marker gene,” as used herein, refers to a nucleic acid sequence that enables cells expressing that sequence to be specifically selected with respect to or against a corresponding selection agent. Suitable selection marker genes are known in the art, for example, in International Publication No. 1992 / 08796 and International Publication No. 1994 / 28143; Wigler et al., Proc.Natl.Acad.Sci.USA, 77:3567 (1980); O'Hare et al., Proc.Natl.Acad.Sci.USA, 78:1527 (1981); Mulligan & Berg, Proc.Natl.Acad.Sci.USA, 78:2072 (1981); Colberre-Garapin et al., J.Mol.Biol., 150:1 (1981); Santerre et al., Gene, 30:147 (1984); Kent et al., Science, 237:901-903 (1987); Wigler et al. This is described in al., Cell, IP.223 (1977); Szybalska & Szybalski, Proc. Natl. Acad. Sci. USA, 48:2026 (1962); Lowy et al., Cell, 22:817 (1980); and U.S. Patent Nos. 5,122,464 and 5,770,359.

[0100] In some embodiments, the vector is an “episome expression vector” or “episome,” which can replicate in host cells and persist as an extrachromosomal segment of DNA within the host cell in the presence of appropriate selective pressure (see, e.g., Conese et al., Gene Therapy, 11:1735-1742 (2004)). Representative commercially available episome expression vectors include, but are not limited to, episome plasmids utilizing Epstein-Barr nuclear antigen 1 (EBNA1) and Epstein-Barr virus (EBV) replication origin (oriP). Vectors pREP4, pCEP4, pREP7, and pcDNA3.1 from Invitrogen (Carlsbad, CA) and pB-CMV from Stratagene (La Jolla, CA) represent non-exclusive examples of episome vectors that use the SV40 replication origin instead of the T-antigen and EBNA1 and oriP.

[0101] In some embodiments, the vector is an "integrated expression vector" that may be randomly integrated into the host cell's DNA or may contain a recombination site that enables recombination between the expression vector and a specific site in the host cell's chromosomal DNA. Such integrated expression vectors may utilize endogenous expression regulatory sequences on the host cell's chromosome that result in the expression of the desired protein. Examples of vectors that integrate in a site-specific manner include components of the cre-lox system, such as the flp-in system from Invitrogen (Carlsbad, CA) (e.g., pcDNA® 5 / FRT) or those found in the pExchange-6 Core vector from Stratagene (La Jolla, CA). Examples of vectors that integrate randomly into the host cell chromosome include, for example, pcDNA3.1 from Invitrogen (Carlsbad, CA) (when introduced in the absence of a T-antigen) and pCI or pFNI OA(ACT)FLEXI® from Promega (Madison, WI).

[0102] In some embodiments, the virus is a viral vector. Typical viral expression vectors include, but are not limited to, adenovirus vectors (e.g., the adenovirus Per.C6 system available from Crucell, Inc. (Leiden, The Netherlands)), lentiviral vectors (e.g., the lentiviral pLP1 from Life Technologies (Carlsbad, CA)), and retroviral vectors (e.g., pFB-ERV plus pCFB-EGSH from Stratagene (La Jolla, CA)). In preferred embodiments, the viral vector is a lentiviral vector.

[0103] A vector comprising the nucleic acid of the present invention encoding a CAR can be introduced into a host cell capable of expressing the thereby encoded CAR, including any suitable prokaryotic or eukaryotic cell. A preferred host cell is one that can be easily and reliably proliferated, has a reasonably fast growth rate, has a well-characterized expression system, and can be easily and efficiently transformed or transfected.

[0104] As used herein, the term “host cell” refers to any type of cell that may contain an expression vector. A host cell may be a eukaryotic cell, e.g., a plant, animal, fungus, or algae, or a prokaryotic cell, e.g., a bacterium or protist. A host cell may be a cultured cell or a primary cell, i.e., it may be isolated directly from an organism, e.g., a human. A host cell may be an adherent cell or a suspension cell, i.e., a cell that grows in a suspension. Suitable host cells are known in the art and include, for example, DH5α, Escherichia coli (E. coli) cells, Chinese hamster ovary cells, monkey VERO cells, COS cells, HEK293 cells, and the like. In a preferred embodiment, the host cell is HEK293 cells. In some embodiments, HEK293 cells are derived from the ATCC SD-3515 strain. In some embodiments, HEK293 cells are derived from the IU-VPF MCB strain. In some embodiments, HEK293 cells are derived from the IU-VPF MWCB strain. In some embodiments, the host cell may be a peripheral blood lymphocyte (PBL), a peripheral blood mononuclear cell (PBMC), or a natural killer (NK) cell. Preferably, the host cell is a natural killer (NK) cell. More preferably, the host cell is a T cell.

[0105] To amplify or replicate recombinant expression vectors, the host cells may be prokaryotic cells, such as DH5α cells. To produce viruses from viral expression vectors, the host cells may be eukaryotic cells, such as HEK293 cells. To produce recombinant CARs, the host cells may be mammalian cells. Preferably, the host cells are human cells. The host cells may be of any cell type, may originate from any tissue type, and may be at any developmental stage. Methods for selecting suitable mammalian host cells, as well as methods for cell transformation, culture, amplification, screening, and purification, are known in the art.

[0106] In some embodiments, this disclosure provides isolated host cells expressing nucleic acid sequences encoding CARs as described herein.

[0107] In some embodiments, the host cell is a T cell. The T cells of this disclosure may be any T cells, such as cultured T cells, e.g., primary T cells, or T cells derived from cultured T cell lines, or T cells obtained from mammals. When obtained from mammals, T cells may be obtained from a number of sources, including but not limited to blood, bone marrow, lymph nodes, thymus, or other tissues or body fluids. T cells may also be enriched or purified. The T cells are preferably human T cells (e.g., isolated from humans). The T cells may be of any developmental stage, including but not limited to CD4+ / CD8+ double-positive T cells, CD4+ helper T cells, e.g., Th and Th2 cells, CD8+ T cells (e.g., cytotoxic T cells), tumor-infiltrating cells, memory T cells, naive T cells, etc. In one embodiment, the T cells are CD8+ T cells or CD4+ T cells. T cell lines are available from, for example, the American Type Culture Collection (ATCC, Manassas, VA) and the German Collection of Microorganisms and Cell Cultures (DSMZ), including Jurkat cells (ATCC TIB-152), Sup-Tl cells (ATCC CRL-1942), RPMI 8402 cells (DSMZ ACC-290), Karpas 45 cells (DSMZ ACC-545), and their derivatives.

[0108] In some embodiments, the host cells are natural killer (NK) cells. NK cells are a type of cytotoxic lymphocyte that plays a role in the innate immune system. Defined as large granular lymphocytes, NK cells constitute a third type of cell differentiated from common lymphoid progenitor cells, from which B and T lymphocytes also arise (see, for example, Immunobiology, 5th ed., Janeway et al., eds., Garland Publishing, New York, NY (2001)). NK cells differentiate and mature in the bone marrow, lymph nodes, spleen, tonsils, and thymus. After maturation, NK cells enter the bloodstream as large lymphocytes with characteristic cytotoxic granules. NK cells can recognize and kill certain abnormal cells, such as certain tumor cells and virus-infected cells, and are considered important in innate immune defense against intracellular pathogens. As described above with respect to T cells, NK cells can be any NK cells, such as cultured NK cells, e.g., primary NK cells, or NK cells derived from cultured NK cells, or NK cells obtained from mammals. When obtained from mammals, NK cells can be obtained from a number of sources, including but not limited to blood, bone marrow, lymph nodes, thymus, or other tissues or body fluids. NK cells can also be enriched or purified. Preferably, NK cells are human NK cells (e.g., isolated from humans). NK cell lines are available, for example, from the American Type Culture Collection (ATCC, Manassas, VA), including, for example, NK-92 cells (ATCC CRL-2407), NK92MI cells (ATCC CRL-2408), and their derivatives.

[0109] In some embodiments, the nucleic acid sequence encoding the CAR may be introduced into a cell by “transfection,” “transformation,” or “transduction.” “Transfection,” “transformation,” or “transduction,” as used herein, refers to the introduction of one or more exogenous polynucleotides into a host cell by using physical or chemical methods. Many transfection techniques are known in this art, including, for example, calcium phosphate DNA coprecipitation (see, e.g., Murray EJ (ed.), Methods in Molecular Biology, Vol. 7, Gene Transfer and Expression Protocols, Humana Press (1991)); DEAE-dextran; electroporation; cationic liposome-mediated transfection; tungsten particle-enhanced microparticle gun (Johnston, Nature, 346:776-777 (1990)); and strontium phosphate DNA coprecipitation (Brash et al., Mol. Cell Biol., 7:2031-2034 (1987)). Phages or viral vectors can be introduced into host cells after proliferation of infectious particles in suitable packaging cells, many of which are commercially available.

[0110] Chimeric antigen receptor International Publication No. 2018 / 028647 is incorporated herein by reference in its entirety. U.S. Patent Application Publication No. 2018 / 0230225 is also incorporated herein by reference in its entirety. Both publications describe BCMA-targeting chimeric antigen receptors (CARs) that are useful in this disclosure.

[0111] This disclosure provides a method for treating a target with cells expressing a chimeric antigen receptor (CAR). A CAR comprises an extracellular antigen-binding domain containing one or more single-domain antibodies. In various embodiments and settings, a BCMA-targeting CAR (also referred to herein as a "BCMA CAR") is provided, comprising a polypeptide including (a) an extracellular antigen-binding domain containing an anti-BCMA binding moiety; (b) a transmembrane domain; and (c) an intracellular signaling domain. In some embodiments, the anti-BCMA binding moiety is camelid, chimeric, human, or humanized. In some embodiments, the intracellular signaling domain comprises a primary intracellular signaling domain of an immune effector cell (such as a T cell). In some embodiments, the primary intracellular signaling domain is derived from CD4. In some embodiments, the primary intracellular signaling domain is derived from CD3-ζ. In some embodiments, the intracellular signaling domain comprises a co-stimulatory signaling domain. In some embodiments, the co-stimulatory signaling domain is derived from a co-stimulatory molecule selected from the group consisting of ligands and combinations thereof of CD27, CD28, CD137, OX40, CD30, CD40, CD3, LFA-1, ICOS, CD2, CD7, LIGHT, NKG2C, B7-H3, CD83. In certain embodiments, the transmembrane domain is derived from CD137.

[0112] In some embodiments, the BCMA CAR further comprises a hinge domain (such as a CD8-α hinge domain) located between the C-terminus of the extracellular antigen-binding domain and the N-terminus of the transmembrane domain. In some embodiments, the BCMA CAR further comprises a signal peptide (such as a CD8-α signal peptide) located at the N-terminus of the polypeptide. In some embodiments, the polypeptide comprises, from N-terminus to C-terminus, a CD8-α signal peptide, an extracellular antigen-binding domain, a CD8-α hinge domain, a CD28 transmembrane domain, a first co-stimulatory signaling domain derived from CD28, a second co-stimulatory signaling domain derived from CD137, and a primary intracellular signaling domain derived from CD4. In some embodiments, the polypeptide comprises, from N-terminus to C-terminus, a CD8-α signal peptide, an extracellular antigen-binding domain, a CD8-α hinge domain, a CD8-α transmembrane domain, a second co-stimulatory signaling domain derived from CD137, and a primary intracellular signaling domain derived from CD3-ζ. In some embodiments, the BCMA CAR is monospecific. In some embodiments, BCMA CAR is monovalent.

[0113] The present invention also provides CARs having two or more (but not limited to two, three, four, five, six, or more) binding sites that specifically bind to an antigen, such as BCMA. In some embodiments, one or more binding sites are antigen-binding fragments. In some embodiments, one or more binding sites include a single-domain antibody. In some embodiments, one or more binding sites include a VHH.

[0114] In some embodiments, the CAR is a polyvalent (such as divalent, trivalent, or more valencies) CAR comprising a polypeptide including (a) an extracellular antigen-binding domain having multiple binding sites (such as at least two, three, four, five, six, or more) that specifically bind to an antigen (such as a tumor antigen); (b) a transmembrane domain; and (c) an intracellular signaling domain.

[0115] In some embodiments, the binding site, such as a VHH (including multiple VHHs, or a first VHH and / or a second VHH), is a camelid, a chimera, a human, or a humanized form. In some embodiments, the binding sites or VHHs are linked to each other by peptide bonds or peptide linkers. In some embodiments, each peptide linker is about 50 amino acids long or less (such as one of approximately 35, 25, 20, 15, 10, or 5 amino acids long or less).

[0116] In some embodiments, the first BCMA binding portion and / or the second BCMA binding portion is an anti-BCMA VHH. In some embodiments, the first BCMA binding portion is a first anti-BCMA VHH, and the second BCMA binding portion is a second anti-BCMA VHH.

[0117] In some embodiments, the first BCMA-binding portion and the second BCMA-binding portion are linked to each other by a peptide linker. In some embodiments, the peptide linker comprises the amino acid sequence of SEQ ID NO: 3. In some embodiments, the peptide linker comprises a polypeptide encoded by the nucleic acid sequence of SEQ ID NO: 11.

[0118] In some embodiments, the CAR further comprises a hinge domain (such as a CD8-α hinge domain) located between the C-terminus of the extracellular antigen-binding domain and the N-terminus of the transmembrane domain. In some embodiments, the CAR further comprises a signal peptide (such as a CD8-α signal peptide) located at the N-terminal end of the polypeptide.

[0119] While we do not wish to be constrained by theory, polyvalent CARs, or such CARs containing an extracellular antigen-binding domain comprising a first BCMA-binding site and a second BCMA-binding site, may be particularly suitable for targeting multimeric antigens by synergistic binding by different antigen-binding sites, or for enhancing binding affinity or avidity to antigens. Improved avidity may result in an increase of 4.0 × 10⁶ per kilogram of body weight of the subject. 4 ~1.0×106 individual CAR-T cells, or 3.0×10 6 ~1.0×10 8 doses in the range such as the number of total CAR-T expressing cells, it may be possible to substantially reduce the dose of CAR-T cells necessary to achieve a therapeutic effect. Monovalent CARs, such as bb2121, may need to be administered at 5 to 10 times such amounts to achieve equivalent effects. In various embodiments, the reduction in the dosage range can result in a substantial reduction in cytokine release syndrome (CRS) and other potentially dangerous side effects of CAR-T therapy.

[0120] The various binding portions (e.g., the extracellular antigen-binding domain comprising the first BCMA-binding portion and the second BCMA-binding portion) in the CARs described herein may be connected to each other via a peptide linker. The peptide linkers connecting different binding portions (such as VHHs) may be the same or different. Different domains of the CAR may also be connected to each other via a peptide linker. In some embodiments, the binding portions (such as VHHs) are directly connected to each other without any peptide linker.

[0121] The peptide linkers of the CARs described herein may be of any preferred length. In some embodiments, the peptide linker is at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 50, 75, 100 amino acid lengths or more. In some embodiments, the peptide linker does not exceed about 100, 75, 50, 40, 35, 30, 25, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5 amino acid lengths or less. In some embodiments, the length of the peptide linker is one of the following: approximately 1 amino acid to approximately 10 amino acids, approximately 1 amino acid to approximately 20 amino acids, approximately 1 amino acid to approximately 30 amino acids, approximately 5 amino acids to approximately 15 amino acids, approximately 10 amino acids to approximately 25 amino acids, approximately 5 amino acids to approximately 30 amino acids, approximately 10 amino acids to approximately 30 amino acids, approximately 30 amino acids to approximately 50 amino acids, approximately 50 amino acids to approximately 100 amino acids, or approximately 1 amino acid to approximately 100 amino acids.

[0122] The CAR of this application includes a transmembrane domain which may be directly or indirectly connected to an extracellular antigen-binding domain.

[0123] The CAR may include a T cell activating moiety. The T cell activating moiety may be derived from or obtained from any suitable molecule. In one embodiment, for example, the T cell activating moiety includes a transmembrane domain. The transmembrane domain may be derived from or obtained from any molecule known in the art. For example, the transmembrane domain may be obtained from or derived from the CD8a molecule or the CD28 molecule. Although we do not wish to be constrained by theory, CD8 is a transmembrane glycoprotein that acts as a co-receptor for T cell receptors (TCRs) and is mainly expressed on the surface of cytotoxic T cells. The most common form of CD8 exists as a dimer composed of a CD8 alpha (CD8α) chain and a CD8 beta (CD8β) chain. CD28 is expressed on T cells and provides the costimulatory signals necessary for T cell activation. CD28 is the receptor for CD80 (B7.1) and CD86 (B7.2). In a preferred embodiment, CD8α and CD28 are human.

[0124] In addition to the transmembrane domain, the T cell activation moiety may further include an intracellular (i.e., cytoplasmic) T cell signaling domain. The intercellular T cell signaling domain may be obtained from or derived from the CD28 molecule, the CD3 zeta (ζ) molecule or modified versions thereof, the human Fc receptor gamma (FcRγ) chain, the CD27 molecule, the OX40 molecule, the 4-1BB molecule, or other intracellular signaling molecules known in the art. While we do not wish to be constrained by theory, (1) CD28 is an important T cell marker for T cell costimulation; (2) CD3ζ associates with the TCR to generate a signal and possesses an immunoreceptor tyrosine activation motif (ITAM); and (3) 4-1BB, also known as CD137, transmits potent costimulatory signals to T cells, promoting T lymphocyte differentiation and enhancing long-term survival. In preferred embodiments, CD28, CD3 zeta, 4-1BB, OX40, and CD27 are human.

[0125] The T cell activation domain of a CAR encoded by the nucleic acid sequences disclosed herein may include any combination of one of the aforementioned transmembrane domains and one or more of the aforementioned intercellular T cell signaling domains. For example, the nucleic acid sequences disclosed herein may encode a CAR comprising a CD28 transmembrane domain and intracellular T cell signaling domains of CD28 and CD3 zeta. Alternatively, for example, the nucleic acid sequences disclosed herein may encode a CAR comprising a CD8α transmembrane domain and intracellular T cell signaling domains of CD28, CD3 zeta, Fc receptor gamma (FcRγ) chain, and / or 4-1BB.

[0126] In some embodiments, the CAR polypeptide further comprises a signal peptide located at the N-terminus of the polypeptide. In some embodiments, the signal peptide is derived from CD8-alpha. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 1. In some embodiments, the signal peptide comprises a polypeptide encoded by the nucleic acid sequence of SEQ ID NO: 9.

[0127] In certain embodiments, the transmembrane domain comprises the amino acid sequence of SEQ ID NO: 6. In certain embodiments, the transmembrane domain comprises a polypeptide encoded by the nucleic acid sequence of SEQ ID NO: 14.

[0128] In some embodiments, the intracellular signaling domain includes the primary intracellular signaling domain of an immune effector cell. In some embodiments, the intracellular signaling domain is derived from CD3ζ. In some embodiments, the intracellular signaling domain includes at least one co-stimulatory signaling domain. In some embodiments, the intracellular signaling domain includes the amino acid sequence of SEQ ID NO: 8. In some embodiments, the intracellular signaling domain includes a polypeptide encoded by the nucleic acid sequence of SEQ ID NO: 16. In some embodiments, the intracellular signaling domain includes the amino acid sequence of SEQ ID NO: 7. In some embodiments, the intracellular signaling domain includes a polypeptide encoded by the nucleic acid sequence of SEQ ID NO: 15.

[0129] In some embodiments, the CAR polypeptide further comprises a hinge domain located between the C-terminus of the extracellular antigen-binding domain and the N-terminus of the transmembrane domain. In some embodiments, the hinge domain comprises the amino acid sequence of SEQ ID NO: 5. In some embodiments, the hinge domain comprises a polypeptide encoded by the nucleic acid sequence of SEQ ID NO: 13.

[0130] In some embodiments, CAR includes one or more of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, and 8, or all of them. In one embodiment, CAR includes SEQ ID NOs: 17. In some embodiments, CAR includes a polypeptide encoded by one or more of SEQ ID NOs: 9, 10, 11, 12, 13, 14, 15, and 16, or all of them.

[0131] In a preferred embodiment, the CAR includes a first VHH domain comprising CDR1, CDR2, and CDR3 of the VHH domain containing the amino acid sequence of SEQ ID NO: 2, and a second VHH domain comprising CDR1, CDR2, and CDR3 of the VHH domain containing the amino acid sequence of SEQ ID NO: 4. In a preferred embodiment, the first VHH domain is linked to the second VHH domain by a linker containing the amino acid sequence of SEQ ID NO: 3. In a particularly preferred embodiment, the first VHH domain comprises CDR1 containing the amino acid sequence of SEQ ID NO: 18, CDR2 containing the amino acid sequence of SEQ ID NO: 19, and CDR3 containing the amino acid sequence of SEQ ID NO: 20, and the second VHH domain comprises CDR1 containing the amino acid sequence of SEQ ID NO: 21, CDR2 containing the amino acid sequence of SEQ ID NO: 22, and CDR3 containing the amino acid sequence of SEQ ID NO: 23. In a further preferred embodiment, the CAR includes a first VHH domain containing the amino acid sequence of SEQ ID NO: 2 and a second VHH domain containing the amino acid sequence of SEQ ID NO: 4.

[0132] Immunoeffector cell composition "Immune effector cells" are immune cells capable of performing immune effector functions. In some embodiments, immune effector cells express at least FcγRIII and perform ADCC effector functions. Examples of ADCC-mediated immune effector cells include peripheral blood mononuclear cells (PBMCs), natural killer (NK) cells, monocytes, cytotoxic T cells, neutrophils, and eosinophils. In some embodiments, immune effector cells are T cells. In some embodiments, T cells are autologous T cells. In some embodiments, T cells are allogeneic T cells. In some embodiments, T cells are CD4+ / CD8-, CD4- / CD8+, CD4+ / CD8+, CD4- / CD8-, or combinations thereof. In some embodiments, T cells produce IL-2, TFN, and / or TNF in conjunction with CAR expression and binding to target cells, such as CD20+ or ​​CD19+ tumor cells. In some embodiments, CD8+ T cells lyse antigen-specific target cells upon CAR expression and binding to target cells.

[0133] Biological methods for introducing vectors into immune effector cells include the use of DNA and RNA vectors. Viral vectors are the most widely used method for inserting genes into mammalian cells, such as human cells. Chemical methods for introducing vectors into immune effector cells include colloidal dispersion systems, such as macromolecular complexes, nanocapsules, microspheres, beads, and lipid-based systems including oil-in-water emulsions, micelles, mixed micelles, and liposomes. An exemplary colloidal system used as a delivery medium in vitro is liposomes (e.g., artificial membrane vesicles).

[0134] This specification includes a CAR comprising a polypeptide comprising (a) an extracellular antigen-binding domain including a first BCMA-binding moiety that specifically binds to a first epitope of BCMA and a second BCMA-binding moiety that specifically binds to a second epitope of BCMA; (b) a transmembrane domain; and (c) an intracellular signaling domain, totaling 3.0 × 10 7 ~1.0×10 8 A drug delivery form is provided that contains 3.0 × 10¹ CAR-T cells, which differ from the first and second epitopes. In a particular embodiment, the drug delivery form contains 3.0 × 10¹ CAR-T cells. 7 ~4.0×10 7 Contains 10 CAR-T cells. In a particular embodiment, the dosage form is 3.5 × 10 7 ~4.5×10 7 Contains CAR-T cells. In a particular embodiment, the dosage form is 4.0 × 10 7 ~5.0×10 7 Contains CAR-T cells. In a particular embodiment, the dosage form is 4.5 × 10 7 ~5.5×10 7 Contains 5.0 × 10⁶ CAR-T cells. In a particular embodiment, the dosage form is 5.0 × 10⁶. 7 ~6.0×10 7 Contains 5.5 × 10¹ CAR-T cells. In a particular embodiment, the dosage form is 5.5 × 10¹⁶ 7 ~6.5×10 7Contains CAR-T cells. In a particular embodiment, the dosage form is 6.0 × 10 7 ~7.0×10 7 Contains CAR-T cells. In a particular embodiment, the dosage form is 6.5 × 10 7 ~7.5×10 7 Contains CAR-T cells. In a particular embodiment, the dosage form is 7.0 × 10 7 ~8.0×10 7 Contains CAR-T cells. In a particular embodiment, the dosage form is 7.5 × 10 7 ~8.5×10 7 Contains CAR-T cells. In a particular embodiment, the dosage form is 8.0 × 10 7 ~9.0×10 7 Contains CAR-T cells. In a particular embodiment, the dosage form is 8.5 × 10 7 ~9.5×10 7 Contains CAR-T cells. In a particular embodiment, the dosage form is 9.0 × 10 7 ~1.0×10 8 Contains 1 CAR-T cell.

[0135] In some embodiments, a CAR comprising (a) an extracellular antigen-binding domain including a first anti-BCMA VHH that specifically binds to a first epitope of BCMA and a second anti-BCMA VHH that specifically binds to a second epitope of BCMA; (b) a transmembrane domain; and (c) an intracellular signaling domain, comprising a polypeptide. 7 ~1.0×10 8 A drug delivery form is provided that includes individual manipulated immune effector cells (such as T cells), which differ from the first and second epitopes. In a particular embodiment, the drug delivery form is 3.0 × 10⁶ 7 ~4.0×10 7 Contains 10 CAR-T cells. In a particular embodiment, the dosage form is 3.5 × 10 7 ~4.5×10 7 Contains CAR-T cells. In a particular embodiment, the dosage form is 4.0 × 10 7 ~5.0×10 7Contains CAR-T cells. In a particular embodiment, the dosage form is 4.5 × 10 7 ~5.5×10 7 Contains 5.0 × 10⁶ CAR-T cells. In a particular embodiment, the dosage form is 5.0 × 10⁶. 7 ~6.0×10 7 Contains 5.5 × 10¹ CAR-T cells. In a particular embodiment, the dosage form is 5.5 × 10¹⁶ 7 ~6.5×10 7 Contains CAR-T cells. In a particular embodiment, the dosage form is 6.0 × 10 7 ~7.0×10 7 Contains CAR-T cells. In a particular embodiment, the dosage form is 6.5 × 10 7 ~7.5×10 7 Contains CAR-T cells. In a particular embodiment, the dosage form is 7.0 × 10 7 ~8.0×10 7 Contains CAR-T cells. In a particular embodiment, the dosage form is 7.5 × 10 7 ~8.5×10 7 Contains CAR-T cells. In a particular embodiment, the dosage form is 8.0 × 10 7 ~9.0×10 7 Contains CAR-T cells. In a particular embodiment, the dosage form is 8.5 × 10 7 ~9.5×10 7 Contains CAR-T cells. In a particular embodiment, the dosage form is 9.0 × 10 7 ~1.0×10 8 Contains 1 CAR-T cell.

[0136] In some embodiments, the cell populations of the CAR-T drug delivery forms described herein include, for example, T cells or T cell populations at various stages of differentiation. T cell differentiation stages include, from minimal to most advanced differentiation, naive T cells, central memory T stem cells, central memory T cells, effector memory T cells, and terminal effector T cells. After antigen exposure, naive T cells proliferate and differentiate into memory T cells, such as central memory T stem cells and central memory T cells, which then differentiate into effector memory T cells. Upon receiving appropriate T cell receptors, co-stimulation, and inflammatory signals, memory T cells further differentiate into terminal effector T cells. See, for example, Restifo.Blood.124.4(2014):476-77; and Joshi et al.J.Immunol.180.3(2008):1309-15.

[0137] Naive T cells may have the following cell surface marker expression patterns: CCR7+, CD62L+, CD45RO-, CD95-. Central memory T stem cells (Tscm) may have the following cell surface marker expression patterns: CCR7+, CD62L+, CD45RO-, CD95+. Central memory T cells (Tcm) may have the following cell surface marker expression patterns: CCR7+, CD62L+, CD45RO+, CD95+. Effector memory T cells (Tem) may have the following cell surface marker expression patterns: CCR7-, CD62L-, CD45RO+, CD95+. Terminally differentiated effector T cells (Teff) may have the following cell surface marker expression patterns: CCR7-, CD62L-, CD45RO-, CD95+. For example, see Gattinoni et al. Nat. Med. 17 (2011): 1290-7; and Flynn et al. Clin. Translat. Immunol. 3 (2014): e20.

[0138] Pharmaceutical compositions and preparations The present application further provides a pharmaceutical composition comprising one of the engineered immunoeffector cells (e.g., BCMA CAR) containing any one of the anti-BCMA antibodies of the present disclosure or any one of the CARs as described herein, and a pharmaceutically acceptable carrier. The pharmaceutical composition may be prepared by mixing any of the immunoeffector cells described herein, having a desired degree of purity, with an optional pharmaceutically acceptable carrier, excipient, or stabilizer (Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)) in the form of a lyophilized formulation or an aqueous solution. In certain embodiments, the CAR-T cell pharmaceutical composition further comprises an excipient selected from dimethyl sulfoxide (DMSO) or dextran-40. In some embodiments, the formulation provided herein contains 5% DMSO.

[0139] The compositions described herein may be administered as part of a pharmaceutical composition comprising one or more carriers. The choice of carrier will be partially determined by a specific nucleic acid sequence, vector, or host cell expressing the CAR disclosed herein, and by a specific method used to administer the nucleic acid sequence, vector, or host cell expressing the CAR disclosed herein. Accordingly, various suitable formulations of the pharmaceutical compositions disclosed herein exist.

[0140] For example, a pharmaceutical composition may contain a preservative. Suitable preservatives may include, for example, methylparaben, propylparaben, sodium benzoate, and benzalkonium chloride. A mixture of two or more preservatives may be used at the discretion of the preservative. The preservative or mixture thereof is typically present in an amount of about 0.0001% to about 2% by weight of the total composition.

[0141] In addition, buffering agents may be used in the composition. Suitable buffering agents include, for example, citric acid, sodium citrate, phosphoric acid, potassium phosphate, and various other acids and salts. Mixtures of two or more buffering agents may be used at the option of choice. The buffering agent or mixture thereof is typically present in an amount of about 0.001% to about 4% by weight of the total composition.

[0142] Compositions comprising nucleic acid sequences encoding the CARs disclosed herein, or host cells expressing the CARs disclosed herein, may be formulated as inclusion complexes such as cyclodextrin inclusion complexes, or as liposomes. Liposomes may act to target host cells (e.g., T cells or NK cells) or the nucleic acid sequences disclosed herein to specific tissues. Liposomes may also be used to increase the half-life of the nucleic acid sequences disclosed herein. Numerous methods are available for preparing liposomes, such as those described in, for example, Szoka et al., Ann. Rev. Biophys. Bioeng., 9:467 (1980), and U.S. Patents No. 4,235,871; No. 4,501,728; No. 4,837,028; and No. 5,019,369. The compositions may utilize time-release, delayed-release, and sustained-release delivery systems so that the delivery of the compositions disclosed herein occurs before sensitization of the site to be treated and with sufficient time to induce sensitization. Many types of delivery systems are available and known to those skilled in the art. Such systems can avoid repeated administration of the compositions, thereby increasing convenience for subjects and physicians, and may be particularly suitable for certain compositional forms and embodiments of the present disclosure.

[0143] In a particular embodiment, CAR-T cells are approximately 1.0 × 10⁶ 5 ~2.0×10 5 cells / kg, 1.5×10 5 ~2.5×10 5 cells / kg, 2.0×10 5 ~3.0×10 5 cells / kg, 2.5×10 5 ~3.5×105 cells / kg, 3.0×10 5 ~4.0×10 5 cells / kg, 3.5×10 5 ~4.5×10 5 cells / kg, 4.0×10 5 ~5.0×10 5 cells / kg, 4.5×10 5 ~5.5×10 5 cells / kg, 5.0×10 5 ~6.0×10 5 cells / kg, 5.5×10 5 ~6.5×10 5 cells / kg, 6.0×10 5 ~7.0×10 5 cells / kg, 6.5×10 5 ~7.5×10 5 cells / kg, 7.0×10 5 ~8.0×10 5 cells / kg, 7.5×10 5 ~8.5×10 5 cells / kg, 8.0×10 5 ~9.0×10 5 cells / kg, 8.5×10 5 ~9.5×10 5 cells / kg, or 9.0×10 5 [[ID=X]]~1.0×10 6 cells / kg and formulated. In a preferred embodiment, the dose is about 0.75×10 6 cells / kg and formulated. In certain embodiments, the CAR-T cells are formulated at a dose of less than 1.0×10 8 cells per subject. Preferably, the dose is administered as a single infusion.

[0144] Methods of Treatment and Use The present application further relates to methods and compositions for use in cellular immunotherapy.

[0145] Note: In the original text, there seems to be a missing closing tag for the range in line . I assumed it should be something like "~1.0×10" and added an "X" tag for clarity in the translation. If this is incorrect, please provide the correct information.In some embodiments, this specification provides a method for treating a target cancer having multiple myeloma, having received one to three prior lines of treatment, including immunomodulatory drug (IMiD) therapy, and being resistant to IMiD, using compositions provided herein. In some embodiments, this specification provides compositions for use in the treatment of a target cancer having multiple myeloma, having received one to three prior lines of treatment, including immunomodulatory drug (IMiD) therapy, and being resistant to IMiD. In some embodiments, the subject has received one prior line of treatment. In some embodiments, the subject has received two prior lines of treatment. In some embodiments, the subject has received three prior lines of treatment.

[0146] In a preferred embodiment, the subject has previously received treatment with IMiD as part of one or more of one to three prior treatment lines. In some embodiments, the IMiD is lenalidomide. In a preferred embodiment, the patient is lenalidomide resistant. In some embodiments, the prior treatment includes pomalidomide. In some embodiments, the IMiD prior treatment line includes a combination of lenalidomide and pomalidomide. In some embodiments, the subject has previously received treatment with a proteasome inhibitor as part of one or more of one to three prior treatment lines. In some embodiments, the proteasome inhibitor is bortezomib, carfilzomib, ixazomib, or any combination thereof. In some embodiments, the subject has previously received treatment with an anti-CD38 antibody as part of one or more of one to three prior treatment lines. In some embodiments, the anti-CD38 antibody is daratumumab and / or isatuximab. In some embodiments, prior treatment includes an IMiD (e.g., lenalidomide), a proteasome inhibitor, and an anti-CD38 antibody (i.e., three prior treatment lines). In some embodiments, the subject has received one prior treatment line including lenalidomide, is lenalidomide resistant, and has optionally received one or two further treatment lines. In some embodiments, the subject has received at least one prior treatment line including lenalidomide and a proteasome inhibitor, and has optionally received one or two further treatment lines.

[0147] This therapy can be optionally used to treat subjects with high-risk features, including, for example, cytogenetic abnormalities, International Staging (ISS) stage III, and / or soft tissue plasmacytoma. In some embodiments, the high-risk feature is a cytogenetic abnormality. In some embodiments, the cytogenetic abnormality is a high-risk cytogenetic abnormality. In some embodiments, the subject has one or more high-risk cytogenetic abnormalities selected from the group including Gain / amp(1q), del(17p), t(4;14), t(14;16), or any combination thereof. In some embodiments, the cytogenetic abnormality includes Gain / amp(1q). In some embodiments, the cytogenetic abnormality includes del(17p). In some embodiments, the cytogenetic abnormality includes t(4;14). In some embodiments, the cytogenetic abnormality includes t(14;16). t(4;14) and t(14;16) are translocations, which are the exchange of portions of chromosomes. del(17p) indicates the loss of a portion of the short arm of chromosome 17. Gain / amp(1q) indicates the gain (e.g., a total of 3 copies) or amplification (e.g., less than a total of 3 copies) of a portion of the long arm of chromosome 1. In some embodiments, the subject has at least two cytogenetic abnormalities. In other embodiments, the subject has two, three, four, five, or more cytogenetic abnormalities. In other embodiments, the cytogenetic abnormalities are standard-risk cytogenetic abnormalities. In some embodiments, the high-risk feature is International Staging Classification (ISS) stage III. In some embodiments, the high-risk feature is soft tissue plasmacytoma.

[0148] In other embodiments, the method provided herein comprises first determining whether a subject has a high-risk feature, wherein the high-risk feature is a cytogenetic abnormality, International Staging (ISS) stage III, and / or soft tissue plasmacytoma; and second administering the composition provided herein to the subject determined to have the high-risk feature. In some embodiments, the subject has multiple myeloma, has received one to three prior lines of treatment, including immunomodulatory drug (IMiD) therapy, and is resistant to IMiD. In some embodiments, the IMiD is lenalidomide. In some embodiments, the high-risk feature is a cytogenetic abnormality. In some embodiments, the cytogenetic abnormality is a high-risk cytogenetic abnormality. In some embodiments, the subject has one or more high-risk cytogenetic abnormalities selected from the group including Gain / amp(1q), del(17p), t(4;14), t(14;16), or any combination thereof. In some embodiments, the cytogenetic abnormality includes Gain / amp(1q). In some embodiments, the cytogenetic abnormality includes del(17p). In some embodiments, the cytogenetic abnormality includes t(4;14). In some embodiments, the cytogenetic abnormality includes t(14;16). In some embodiments, the subject has at least two cytogenetic abnormalities. In other embodiments, the subject has two, three, four, five, or more cytogenetic abnormalities. In other embodiments, the cytogenetic abnormalities are standard-risk cytogenetic abnormalities. In some embodiments, the high-risk feature is International Staging Classification (ISS) stage III. In some embodiments, the high-risk feature is soft tissue plasmacytoma. In some embodiments, the subject has received one prior line of treatment. In some embodiments, the subject has received two prior lines of treatment. In some embodiments, the subject has received three prior lines of treatment. In some embodiments, the prior treatment includes pomalidomide. In some embodiments, prior treatment further includes a proteasome inhibitor, which is optionally bortezomib, carfilzomib, ixazomib, or any combination thereof.In some embodiments, the prior treatment further comprises an anti-CD38 antibody, which is optionally daratumumab and / or isatuximab. In some embodiments, the prior treatment comprises an IMiD (e.g., lenalidomide), a proteasome inhibitor, and an anti-CD38 antibody.

[0149] In further embodiments, the Specified herein provides a method for selectively treating a subject with a composition provided herein, comprising administering the composition to a subject determined to have high-risk features, such as a cytogenetic abnormality, International Staging (ISS) stage III, and / or soft tissue plasmacytoma. In some embodiments, the composition provided herein is intended for use in the treatment of a subject determined to have high-risk features, such as a cytogenetic abnormality, International Staging (ISS) stage III, and / or soft tissue plasmacytoma. In some embodiments, the subject has multiple myeloma, has received one to three prior lines of treatment, including immunomodulatory drug (IMiD) therapy, and is resistant to IMiD. In some embodiments, the IMiD is lenalidomide. In some embodiments, the high-risk feature is a cytogenetic abnormality. In some embodiments, the cytogenetic abnormality is a high-risk cytogenetic abnormality. In some embodiments, the subject has one or more high-risk cytogenetic abnormalities selected from the group including Gain / amp(1q), del(17p), t(4;14), t(14;16), or any combination thereof. In some embodiments, the cytogenetic abnormality includes Gain / amp(1q). In some embodiments, the cytogenetic abnormality includes del(17p). In some embodiments, the cytogenetic abnormality includes t(4;14). In some embodiments, the cytogenetic abnormality includes t(14;16). In some embodiments, the subject has at least two cytogenetic abnormalities. In other embodiments, the subject has two, three, four, five, or more cytogenetic abnormalities. In other embodiments, the cytogenetic abnormalities are standard-risk cytogenetic abnormalities. In some embodiments, the high-risk feature is International Staging System (ISS) stage III. In some embodiments, the high-risk feature is soft tissue plasmacytoma. In some embodiments, the subject has received one prior line of treatment. In some embodiments, the subject has received two prior treatment lines. In some embodiments, the subject has received three prior treatment lines. In some embodiments, the prior treatment includes pomalidomide.In some embodiments, the prior treatment further comprises a proteasome inhibitor, optionally being bortezomib, carfilzomib, ixazomib, or any combination thereof. In some embodiments, the prior treatment further comprises an anti-CD38 antibody, optionally being daratumumab and / or isatuximab. In some embodiments, the prior treatment comprises an IMiD (e.g., lenalidomide), a proteasome inhibitor, and an anti-CD38 antibody.

[0150] In some embodiments of the various methods or uses provided herein, the subject has received further bridging therapy, which is optional and may be at the physician's discretion. Examples of bridging therapy include pomalidomide, bortezomib, dexamethasone, daratumumab, or any combination thereof. Examples include pomalidomide, bortezomib, and dexamethasone. Another exemplary bridging therapy includes daratumumab, pomalidomide, and dexamethasone. In some embodiments, the subject has received bridging therapy approximately every 20 to 30 days, for example, every 21 days or every 28 days. In some embodiments, the subject has received at least one, two, three, four, or more bridging therapies.

[0151] In certain embodiments, the bridging therapy comprises a 28-day cycle including daratumumab on days 1, 8, 15, and 22, pomalidomide for 21 days, and dexamethasone on days 1, 8, 15, and 22. For example, the bridging therapy may include 1800 mg daratumumab on days 1, 8, 15, and 22; 4 mg / day pomalidomide for 21 days; and 40 mg dexamethasone on days 1, 8, 15, and 22. In the embodiments described above, daratumumab may be administered subcutaneously, pomalidomide may be administered orally, and dexamethasone may be administered orally or intravenously.

[0152] In other embodiments, bridging therapy comprises a 21-day cycle including bortezomib on days 1, 4, 8, and 11, pomalidomide for 14 days, and dexamethasone on days 1, 2, 4, 5, 8, 9, 11, and 12. For example, bridging therapy may include 1.3 mg / m² on days 1, 4, 8, and 11. 2 The regimen may include bortezomib; 4 mg / day of pomalidomide for 14 days; and 20 mg of dexamethasone on bridging days 1, 2, 4, 5, 8, 9, 11, and 12. In the above embodiment, bortezomib may be administered subcutaneously, pomalidomide may be administered orally, and dexamethasone may be administered orally.

[0153] In some embodiments, the subject has received lymphocyte depletion therapy after, for example, bridging therapy as disclosed herein. In some embodiments, lymphocyte depletion therapy comprises daily cyclophosphamide and / or fludarabine. In some embodiments, lymphocyte depletion therapy comprises daily cyclophosphamide and fludarabine. In some embodiments, lymphocyte depletion therapy comprises approximately 300 mg / m² daily for 3 days. 2 The concentration of cyclophosphamide is approximately 30 mg / m². 2 Contains fludarabine at this concentration.

[0154] In some embodiments of the various methods or uses provided herein, the dose of CAR T cells is 0.5 to 1.0 × 10⁶ 6 The cell-to-subject weight is 1 kg. In a preferred embodiment, the dose of CAR T cells is approximately 0.75 × 10⁻⁶ 6 The cell-to-subject weight is 1 kg. In some embodiments, the method involves administering a dose of CAR T cells approximately 5 to 7 days after the initiation of lymphocyte depletion therapy. Preferably, the dose is administered as a single infusion. In some embodiments, 0.75 × 10⁶ cells are administered 5 to 7 days after the initiation of lymphocyte depletion. 6 A single intravenous infusion of CAR T cells per kg is administered.

[0155] The anti-BCMA VHH, CAR, and modified immune effector cells (such as CAR-T cells) described herein may be used in methods for treating cancer. In some embodiments, the immune effector cells are autologous. In some embodiments, the immune effector cells are allogeneic.

[0156] In a particular embodiment, CAR-T cells are approximately 1.0 × 10⁶ 5 ~2.0×10 5 cells / kg, 1.5×10 5 ~2.5×10 5 cells / kg, 2.0×10 5 ~3.0×10 5 cells / kg, 2.5×10 5 ~3.5×10 5 cells / kg, 3.0×10 5 ~4.0×10 5 cells / kg, 3.5×10 5 ~4.5×10 5 cells / kg, 4.0×10 5 ~5.0×10 5 cells / kg, 4.5×10 5 ~5.5×10 5 cells / kg, 5.0×10 5 ~6.0×10 5 cells / kg, 5.5×10 5 ~6.5×10 5 cells / kg, 6.0×10 5 ~7.0×10 5 cells / kg, 6.5×10 5 ~7.5×10 5 cells / kg, 7.0×10 5 ~8.0×10 5 cells / kg, 7.5×10 5 ~8.5×10 5 cells / kg, 8.0×10 5 ~9.0×10 5 cells / kg, 8.5×10 5 ~9.5×10 5 cells / kg, 9.0×10 5 ~1.0×10 5 cells / kg, 1.0×10 6 ~2.0×10 6cells / kg, 1.5×10 6 ~2.5×10 6 cells / kg, 2.0×10 6 ~3.0×10 6 cells / kg, 2.5×10 6 ~3.5×10 6 cells / kg, 3.0×10 6 ~4.0×10 6 cells / kg, 3.5×10 6 ~4.5×10 6 cells / kg, 4.0×10 6 ~5.0×10 6 cells / kg, 4.5×10 6 ~5.5×10 6 cells / kg, or 5.0 × 10⁻⁶ 6 ~6.0×10 6 It is administered in doses of cells / kg. In a preferred embodiment, the dose is approximately 0.75 × 10⁻⁶ 6 Contains cells / kg. In certain embodiments, CAR-T cells are approximately 1.0 × 10⁶ per subject. 8 It is administered in cellular doses. Preferably, the dose is administered as a single infusion.

[0157] In a particular embodiment, CAR-T cells are 1.0 × 10⁶ per subject. 8 They are administered in doses less than 10 cells. In certain embodiments, CAR-T cells are approximately 3.0–4.0 × 10⁶ 7 They are administered in cell doses. In certain embodiments, CAR-T cells are administered in doses of approximately 3.5–4.5 × 10⁶ 7 They are administered in cell doses. In certain embodiments, CAR-T cells are approximately 4.0-5.0 × 10⁶ 7 They are administered in cell doses. In certain embodiments, CAR-T cells are approximately 4.5–5.5 × 10⁶ 7 They are administered in cell doses. In certain embodiments, CAR-T cells are approximately 5.0–6.0 × 10⁶ 7 They are administered in cell doses. In certain embodiments, CAR-T cells are approximately 5.5–6.5 × 10⁶ 7 They are administered in cell doses. In certain embodiments, CAR-T cells are approximately 6.0–7.0 × 10⁶ 7They are administered in cell doses. In certain embodiments, CAR-T cells are administered in doses of approximately 6.5–7.5 × 10⁶ 7 They are administered in doses of cells. In certain embodiments, CAR-T cells are approximately 7.0–8.0 × 10⁶ 7 They are administered in cell doses. In certain embodiments, CAR-T cells are approximately 7.5–8.5 × 10⁶ 7 They are administered in cell doses. In certain embodiments, CAR-T cells are approximately 8.0-9.0 × 10⁶ 7 They are administered in cell doses. In certain embodiments, CAR-T cells are approximately 8.5–9.5 × 10⁶ 7 They are administered in cell doses. In certain embodiments, CAR-T cells are approximately 9.0 × 10⁶ 7 ~1.0×10 8 It is administered in cellular doses.

[0158] In certain embodiments, CAR-T cells are approximately 0.693 × 10⁶ 6 The cells are administered at a dose of 10¹⁴ CAR-positive viable T cells / kg. In certain embodiments, CAR-T cells are approximately 0.52 × 10¹⁴ 6 The cells are administered at a dose of 10⁴ CAR-positive viable T cells / kg. In certain embodiments, CAR-T cells are approximately 0.94 × 10⁴ 6 The cells are administered at a dose of 10⁴ CAR-positive viable T cells / kg. In certain embodiments, CAR-T cells are approximately 0.709 × 10⁴ 6 They are administered at a dose of CAR-positive viable T cells / kg. In certain embodiments, CAR-T cells are approximately 0.51 × 10⁶ 6 The cells are administered at a dose of CAR-positive viable T cells / kg. In certain embodiments, CAR-T cells are approximately 0.95 × 10⁶ 6 It is administered at a dose of CAR-positive viable T cells / kg. In certain embodiments, CAR-T cells are administered in an outpatient setting.

[0159] In some embodiments, the composition containing CAR-T cells administered to the subject further comprises an excipient selected from dimethyl sulfoxide (DMSO) or dextran-40. In some embodiments, the composition contains 5% DMSO.

[0160] In certain embodiments, CAR-T cells (e.g., any of the aforementioned doses) are administered in one or more intravenous infusions. In certain embodiments, the administration of the CAR-T cells is via a single intravenous infusion. In certain embodiments, the single intravenous infusion is administered using a single bag of the CAR-T cells. In certain embodiments, the administration of the single bag of the CAR-T cells is completed between the time the single bag of the CAR-T cells is thawed and 3 hours after the single bag of the CAR-T cells is thawed. In certain embodiments, the single intravenous administration is administered using two bags of the CAR-T cells. In certain embodiments, the administration of each of the two bags of the CAR-T cells is completed between the time the first of the two bags of CAR-T cells is thawed and 3 hours after the first bag of the CAR-T cells is thawed.

[0161] In certain embodiments, the time from initial apheresis to CAR-T cell administration is less than 41, 47, 54, 61, 68, 75, 82, 89, 96, 103, 110, 117, 124, 131, 138, 145, 152, 159, 166, or 167 days. In certain embodiments, the time from initial apheresis to CAR-T cell administration is longer than 41, 47, 54, 61, 68, 75, 82, 89, 96, 103, 110, 117, 124, 131, 138, 145, 152, 159, 166, or 167 days.

[0162] In certain embodiments, a lymphocyte depletion regimen is administered before CAR-T cell administration. In certain embodiments, the lymphocyte depletion regimen includes the administration of cyclophosphamide and / or fludarabine. In certain embodiments, the lymphocyte depletion regimen is administered intravenously. In certain embodiments, the lymphocyte depletion regimen is administered 5 to 7 days before CAR-T cell administration. In certain embodiments, the lymphocyte depletion regimen is administered 2 to 4 days before CAR-T cell administration. In certain embodiments, the lymphocyte depletion regimen includes the intravenous administration of cyclophosphamide and fludarabine 5 to 7 days before CAR-T cell administration. In certain embodiments, the lymphocyte depletion regimen includes the intravenous administration of cyclophosphamide and fludarabine 2 to 4 days before CAR-T cell administration. In certain embodiments, the lymphocyte depletion regimen is administered at a dose of 300 mg / m². 2 It contains cyclophosphamide administered intravenously. In certain embodiments, the lymphocyte depletion regimen is 30 mg / m² 2 It includes fludarabine administered intravenously. In some embodiments, the lymphocyte depletion regimen is carried out daily for three days. If CAR-T cell administration is delayed for more than 14 days, the lymphocyte depletion regimen may be repeated.

[0163] In certain embodiments, the CAR-T cell therapy method further includes treating the subject with respect to cytokine release syndrome (CRS) within 3 days of CAR-T cell administration without a significant decrease in in vivo CAR-T cell expansion. In certain embodiments, the treatment of CRS includes administering an IL-6R inhibitor to the subject. In certain embodiments, the IL-6R inhibitor is an antibody. In certain embodiments, the IL-6 inhibitor inhibits IL-6R by binding to its extracellular domain. In certain embodiments, the IL-6R inhibitor prevents IL-6 from binding to IL-6R. In certain embodiments, the IL-6R inhibitor is tocilizumab. CRS can be identified based on the clinical presentation. In some embodiments, other causes of fever, hypoxia, and hypotension are evaluated and treated. Clinical tests can be used to monitor disseminated intravascular coagulation, hematological parameters, and pulmonary, cardiac, renal, and hepatic function. CRS can be managed according to the recommendations in Table 12. This method may include administering levetiracetam anticonvulsant prophylaxis to patients presenting with CRS. In some embodiments, this method includes monitoring patients presenting with grade 2 or higher CRS (e.g., hypotension unresponsive to fluids, or hypoxia requiring oxygen supplementation) by continuous cardiac telemetry and pulse oximetry. In some embodiments, intensive care unit-level monitoring and supportive care may be used for severe or life-threatening CRS. For CRS resistant to first-line interventions such as tocilizumab or tocilizumab and corticosteroids, this method includes alternating treatment options (i.e., higher doses of corticosteroids, alternative anti-cytokine agents, e.g., anti-IL1 and / or anti-TNFα, anti-T-cell therapy). Resistant CRS is characterized by fever and end-organ toxicity (e.g., hypoxia, hypotension) that do not improve within 12 hours of the first-line intervention or the onset of HLH / MAS.

[0164] In certain embodiments, the CAR-T cell therapy method further includes treating the subject with pre-infusion drug therapy comprising an antipyretic and an antihistamine up to one hour before administration of CAR-T cells. In certain embodiments, the antipyretic comprises either paracetamol or acetaminophen. In certain embodiments, the antipyretic is administered to the subject either orally or intravenously. In certain embodiments, the antipyretic is administered to the subject in a dose of 650 mg to 1000 mg. In certain embodiments, the antihistamine comprises diphenhydramine. In certain embodiments, the antihistamine is administered to the subject either orally or intravenously. In certain embodiments, the antihistamine is administered in a dose of 25 mg to 50 mg, or an equivalent dose. A composition comprising host cells expressing a CAR-coding nucleic acid sequence disclosed herein, or a vector containing a CAR-coding nucleic acid sequence disclosed herein, can be administered to mammals using standard administration techniques, including oral, intravenous, intraperitoneal, subcutaneous, pulmonary, percutaneous, intramuscular, intranasal, buccal, sublingual, or suppository administration. The composition is preferably suited for parenteral administration. The term “parenteral” as used herein includes intravenous, intramuscular, subcutaneous, rectal, vaginal, and intraperitoneal administration. More preferably, the composition is administered to mammals using peripheral systemic delivery by intravenous, intraperitoneal, or subcutaneous injection. Most preferably, the composition is administered by intravenous infusion.

[0165] A composition comprising host cells expressing a CAR coding nucleic acid sequence disclosed herein, or a vector comprising a CAR coding nucleic acid sequence disclosed herein, may be administered to a mammal together with one or more additional therapeutic agents that may be co-administered. "Co-administration" means administering one or more additional therapeutic agents and the composition comprising the host cells or vector disclosed herein at a time sufficiently close to the time so that the CAR disclosed herein can enhance the effect of the one or more additional therapeutic agents, or vice versa. In this regard, the composition comprising the host cells or vector disclosed herein may be administered first, and one or more additional therapeutic agents may be administered second, or vice versa.

[0166] The CAR-expressing cells and at least one additional therapeutic agent described herein may be administered simultaneously or sequentially in the same or separate compositions. For sequential administration, the CAR-expressing cells described herein may be administered first, followed by the additional agent, or the order of administration may be reversed.

[0167] In certain embodiments, a lymphocyte depletion regimen is administered before the administration of CAR-T cells. In certain embodiments, the lymphocyte depletion regimen is administered approximately 2 to 7 days before the administration of the CAR-T cells. In certain embodiments, the lymphocyte depletion regimen is administered intravenously. In certain embodiments, the lymphocyte depletion regimen includes the administration of cyclophosphamide or fludarabine. In certain embodiments, the cyclophosphamide is administered at a dose of 300 mg / m². 2 It is administered intravenously. In certain embodiments, the fludarabine is 30 mg / m². 2 It is administered intravenously.

[0168] In a specific embodiment, 300 mg / m² 2 Cyclophosphamide and 30 mg / m² are administered intravenously. 2 The lymphocyte depletion regimen, which includes fludarabine administered intravenously, precedes the administration of the CAR-T cells by approximately 2 to 7 days.

[0169] In certain embodiments, the subject further receives bridging therapy, the bridging therapy comprising a short-term treatment with at least one bridging therapy between apheresis and the lymphocyte apheresis regimen, the at least one bridging drug having previously yielded a stable disease, minimal response, partial response, best partial response, complete response, or severe complete response outcome for the subject. In certain embodiments, the subject had an increase in tumor volume despite the bridging therapy. In certain embodiments, the subject had an increase in tumor volume of about 25% or more despite the bridging therapy. Suitable bridging therapies include, for example, dexamethasone, daratumumab, bortezomib, cyclophosphamide, and pomalidomide. In some embodiments, the bridging therapy comprises pomalidomide, bortezomib, dexamethasone, daratumumab, or any combination thereof. In some embodiments, the bridging therapy comprises dexamethasone. In some embodiments, the bridging therapy comprises daratumumab. In some embodiments, the bridging therapy includes bortezomib. In some embodiments, the bridging therapy includes cyclophosphamide. In some embodiments, the bridging therapy includes pomalidomide. In some embodiments, the bridging therapy includes pomalidomide, bortezomib, and dexamethasone. In some embodiments, the bridging therapy includes daratumumab, pomalidomide, and dexamethasone. In some embodiments, the subjects have received bridging therapy approximately every 10 to 40 days. In some embodiments, the subjects have received bridging therapy approximately every 20 to 30 days. In some embodiments, the subjects have received bridging therapy approximately every 21 days. In some embodiments, the subjects have received bridging therapy approximately every 15 days. In some embodiments, the subjects have received bridging therapy approximately every 25 days. In some embodiments, the subjects have received bridging therapy approximately every 21 days. In some embodiments, the subjects have received bridging therapy approximately every 28 days. In some embodiments, the subjects had previously received bridging therapy approximately every 30 days.In some embodiments, the subject has received bridging therapy approximately every 35 days. In some embodiments, the subject has received at least one, two, three, four, or more bridging therapies. In some embodiments, the subject has received at least one bridging therapy. In some embodiments, the subject has received at least two bridging therapies. In some embodiments, the subject has received at least three bridging therapies. In some embodiments, the subject has received at least four bridging therapies. In some embodiments, the subject has received at least five bridging therapies. In some embodiments, the subject has received at least six bridging therapies.

[0170] In certain embodiments, the subject is treated with pre-administered drug therapy including an antipyretic and an antihistamine up to approximately one hour before the administration of the CAR-T cells. In certain embodiments, the antipyretic comprises either paracetamol or acetaminophen. In certain embodiments, the antipyretic is administered to the subject either orally or intravenously. In certain embodiments, the antipyretic is administered to the subject in a dose between 650 mg and 1000 mg. In certain embodiments, the antihistamine comprises diphenhydramine. In certain embodiments, the antihistamine is administered to the subject either orally or intravenously. In certain embodiments, the antihistamine is administered in a dose between 25 mg and 50 mg, or an equivalent thereto. In certain embodiments, the antipyretic comprises either paracetamol or acetaminophen, and is administered to the subject orally or intravenously in a dose between 650 mg and 1000 mg; and the antihistamine comprises diphenhydramine, and is administered to the subject orally or intravenously in a dose between 25 mg and 50 mg, or an equivalent thereto.

[0171] In some embodiments, this method involves administering cyclophosphamide 300 mg / m² daily for three days prior to CAR-T cell administration.2 Intravenous (IV) and fludarabine 30 mg / m² 2 This includes administering a lymphocyte depletion chemotherapy regimen including IV, and administering pre-infusion drug therapy including an antipyretic (such as oral or intravenous acetaminophen 650-1000 mg) and an antihistamine (such as oral or intravenous diphenhydramine 25-50 mg or equivalent), where, CAR-T cells are administered 2-4 days after the completion of lymphocyte depletion chemotherapy, and CAR-T cells are administered 30 to 60 minutes after the administration of pre-infusion drug therapy.

[0172] In some embodiments, CAR-T cell administration is withheld or delayed if the patient exhibits any of the following conditions: clinically significant active infection or inflammatory disorder; or non-hematological toxicity of cyclophosphamide and fludarabine conditioning of grade 3 or higher, excluding grade 3 nausea, vomiting, diarrhea, or constipation. CAR-T cell administration must be delayed until these events resolve to grade 1 or lower. In some embodiments, prophylactic systemic corticosteroids are not administered.

[0173] In some embodiments, the method further includes diagnosing the subject with respect to cytokine release syndrome (CRS). In preferred embodiments, the diagnosis is made according to the consensus grading system of the American Society of Transplantation and Cellular Therapy (ASTCT), formerly the American Society for Blood and Marrow Transplantation (ASBMT). A non-limiting overview of the ASTCT consensus grading system for CRS diagnosis is provided in Table 13.

[0174] In some embodiments, the method further includes treating the subject with respect to cytokine release syndrome (CRS). In some embodiments, the treatment of CRS is with an antipyretic. In some embodiments, the treatment of CRS is with anti-cytokine therapy. In some embodiments, the treatment of CRS is performed about three days after infusion. In some embodiments, the treatment of CRS is performed without significantly reducing CAR-T cell expansion in vivo. In certain embodiments, the method further includes treating the subject with respect to cytokine release syndrome about three days after the administration of CAR-T cells without significantly reducing the expansion of CAR-T cells in vivo. In some embodiments, the treatment of CRS includes administering an IL-6R inhibitor to the subject. In some embodiments, the IL-6R inhibitor is an antibody. In some embodiments, the antibody inhibits IL-6R by binding to its extracellular domain. In some embodiments, the IL-6R inhibitor prevents IL-6 from binding to IL-6R. In some embodiments, the IL-6R inhibitor is tocilizumab. In some embodiments, anti-cytokine therapy includes the administration of tocilizumab. In some embodiments, anti-cytokine therapy includes the administration of steroids. In some embodiments, treatment of CRS includes treatment with a monoclonal antibody other than tocilizumab. In some embodiments, the antibody other than tocilizumab targets a cytokine. In some embodiments, the cytokine targeted by the antibody other than tocilizumab is IL-1. In some embodiments, the IL-1 targeted antibody is anakinra. In some embodiments, the cytokine targeted by the antibody other than tocilizumab is TNFα. In some embodiments, treatment of CRS includes administering corticosteroids to the subject. In some embodiments, treatment of CRS includes the use of vasopressors. In some embodiments, treatment of CRS includes intubation or mechanical ventilation. In some embodiments, treatment of CRS includes administering cyclophosphamide to the subject. In some embodiments, treatment of CRS includes administering etanercept to the subject. In some embodiments, treatment of CRS includes administering levetiracetam to the subject. In some embodiments, treatment for CRS includes supportive care.

[0175] In some embodiments, the method further includes diagnosing the subject with respect to immune cell effector-associated neurotoxicity (ICANS). In some embodiments, the diagnosis is made according to the National Cancer Institute Common Terminology Criteria for Adverse Events (NCI CTCAE) criteria. In some embodiments, the diagnosis is made according to the NCI CTCAE criteria, version 5.0. In some embodiments, the diagnosis is made according to the American Society of Transplantation and Cellular Therapy (ASTCT) consensus grading system. In some embodiments, neurotoxicity consistent with ICAN exists. A non-limiting overview of the ASTCT consensus grading system for ICANS diagnosis is provided in Table 14. In some embodiments, treatment of ICANS includes administering an IL-6R inhibitor to the subject. In some embodiments, the IL-6R inhibitor is an antibody. In some embodiments, the antibody inhibits IL-6R by binding to its extracellular domain. In some embodiments, the IL-6R inhibitor prevents IL-6 from binding to IL-6R. In some embodiments, the IL-6R inhibitor is tocilizumab. In some embodiments, treatment of ICANs includes administering an IL-1 inhibitor to the subject. In some embodiments, the IL-1 inhibitor is an antibody. In a preferred embodiment, the IL-1 inhibitory antibody is anakinra. In some embodiments, treatment of ICANs includes administering a corticosteroid to the subject. In some embodiments, treatment of ICANs includes administering levetiracetam to the subject. In some embodiments, treatment of ICANs includes administering dexamethasone to the subject. In some embodiments, treatment of ICANs includes administering methylprednisone sodium succinate to the subject. In some embodiments, treatment of ICANs includes administering pethidine to the subject.In some embodiments, treatment of ICANS involves administering one or more of the following tocilizumab, anakinra, corticosteroids, levetiracetam, dexamethasone, methylprednisone sodium succinate, or pethidine to the target.

[0176] This method is also applicable when neurotoxicity is suspected to occur during CRS, or vice versa. Corticosteroids that follow more aggressive interventions based on CRS and neurotoxicity grades as listed in Tables 1 and 2 of the approved labeling. Tocilizumab according to the CRS grade listed in Table 1 of the approved labeling. • Anticonvulsant drug therapy in accordance with the neurotoxicity listed in Table 2 of the approved labeling. This may include administering [a drug].

[0177] In some embodiments, the method further includes diagnosing the subject with respect to cytopenia. In some embodiments, cytopenia includes one or more or all of lymphopenia, neutropenia, and thrombocytopenia. Although not bound by theory, lymphopenia of grade 3 or grade 4 but not grade 2 or lower is 0.5 × 10 per liter of the subject's blood sample. 9Neutropenia of grade 3 or grade 4 but not grade 2 is characterized by a neutrophil count of less than 1,000 cells per microliter of the subject's blood sample, and thrombocytopenia of grade 3 or grade 4 but not grade 2 is characterized by a platelet count of less than 50,000 cells per microliter of the subject's blood sample. In some embodiments, more than 75% of subjects with grade 3 or grade 4 lymphopenia after CAR-T cell administration recover to grade 2 or less lymphopenia 60 days after CAR-T cell administration. In some embodiments, more than 80% of subjects with grade 3 or grade 4 lymphopenia after CAR-T cell administration recover to grade 2 or less lymphopenia 60 days after CAR-T cell administration. In some embodiments, more than 85% of subjects with grade 3 or grade 4 lymphopenia after CAR-T cell administration recover to grade 2 or less lymphopenia 60 days after CAR-T cell administration. In some embodiments, more than 90% of subjects with grade 3 or grade 4 lymphopenia after CAR-T cell administration recover to grade 2 or lower lymphopenia 60 days after CAR-T cell administration. In some embodiments, more than 70% of subjects with grade 3 or grade 4 neutropenia after CAR-T cell administration recover to grade 2 or lower neutropenia 60 days after CAR-T cell administration. In some embodiments, more than 75% of subjects with grade 3 or grade 4 neutropenia after CAR-T cell administration recover to grade 2 or lower neutropenia 60 days after CAR-T cell administration. In some embodiments, more than 80% of subjects with grade 3 or grade 4 neutropenia after CAR-T cell administration recover to grade 2 or lower neutropenia 60 days after CAR-T cell administration. In some embodiments, more than 85% of subjects with grade 3 or grade 4 neutropenia after CAR-T cell administration recover to grade 2 or lower neutropenia 60 days after CAR-T cell administration.In some embodiments, more than 30% of subjects with grade 3 or grade 4 thrombocytopenia after CAR-T cell administration recover to grade 2 or lower thrombocytopenia 60 days after CAR-T cell administration. In some embodiments, more than 34% of subjects with grade 3 or grade 4 thrombocytopenia after CAR-T cell administration recover to grade 2 or lower thrombocytopenia 60 days after CAR-T cell administration. In some embodiments, more than 38% of subjects with grade 3 or grade 4 thrombocytopenia after CAR-T cell administration recover to grade 2 or lower thrombocytopenia 60 days after CAR-T cell administration. In some embodiments, more than 42% of subjects with grade 3 or grade 4 thrombocytopenia after CAR-T cell administration recover to grade 2 or lower thrombocytopenia 60 days after CAR-T cell administration.

[0178] When a host cell expressing the CAR coding nucleic acid sequence disclosed herein, or a composition comprising a vector containing the CAR coding nucleic acid sequence disclosed herein, is administered to a mammal (e.g., human), the biological activity of the CAR can be measured by any suitable method known in the art. According to the method disclosed herein, the CAR binds to BCMA on multiple myeloma cells, and the multiple myeloma cells are destroyed. The binding of the CAR to BCMA on the surface of multiple myeloma cells can be assayed using any suitable method known in the art, including, for example, ELISA and flow cytometry. The ability of the CAR to destroy multiple myeloma cells can be measured using any suitable method known in the art, such as the cytotoxic assay described in, for example, Kochenderfer et al., J. Immunotherapy, 32(7):689-702 (2009) and Herman et al., J. Immunological Methods, 285(1):25-40 (2004). The biological activity of CARs can also be measured by assaying the expression of specific cytokines such as CD107a, IFNγ, IL-2, and TNF.

[0179] The methods described herein may be used to treat a variety of cancers, including both solid and liquid tumors. In a particular embodiment, the methods are used to treat multiple myeloma. The methods described herein may be used in an adjuvant setting or a neoadjuvant setting as a first therapy, second therapy, third therapy, or combination therapy, together with other types of cancer therapies known in the art, such as chemotherapy, surgery, radiation therapy, gene therapy, immunotherapy, bone marrow transplantation, stem cell transplantation, targeted therapy, cryotherapy, ultrasound therapy, photodynamic therapy, and radiofrequency ablation.

[0180] In certain embodiments, the cancer is multiple myeloma. In certain embodiments, the cancer is stage I, stage II, or stage III, and / or stage A or stage B, according to the Dury-Salmon staging system. In certain embodiments, the cancer is stage I, stage II, or stage III, according to the International Staging System published by the International Myeloma Working Group (IMWG). In some embodiments, the multiple myeloma drug is progressive.

[0181] In certain embodiments, the subject received prior treatment with one or more lines of treatment. In some embodiments, the number of prior treatment lines is 1. In certain embodiments, the number of prior treatment lines is 2. In some embodiments, the number of prior treatment lines is 3. In some embodiments, the number of prior treatment lines is 4. In some embodiments, the number of prior treatment lines is 5. In certain embodiments, the prior treatment lines include surgery, radiotherapy, or autologous or allogeneic transplantation, or any combination of such treatments. In certain embodiments, prior treatment includes treatment with a drug that is a proteasome inhibitor (PI). Non-limiting examples of PIs include bortezomib, carfilzomib, and ixazomib. In certain embodiments, prior treatment includes treatment with a drug that is an immunomodulatory agent (IMiD). Non-limiting examples of IMiDs include lenalidomide, pomalidomide, and thalidomide. In preferred embodiments, prior treatment includes treatment with lenalidomide, or optionally treatment with lenalidomide and a proteasome inhibitor. In a preferred embodiment, subjects received at least one prior treatment line, optionally including lenalidomide and a proteasome inhibitor, and optionally one or two further prior treatment lines. In a particular embodiment, prior treatment includes treatment with a drug that is a corticosteroid. Non-limiting examples of corticosteroids include dexamethasone and prednisone. In a particular embodiment, prior treatment includes treatment with a drug that is an alkylating agent. In a particular embodiment, prior treatment includes treatment with a drug that is an anthracycline. In a particular embodiment, prior treatment includes treatment with a drug that is an anti-CD38 antibody. Non-limiting examples of anti-CD38 antibodies include daratumumab, isatuximab, and the investigational antibody TAK-079. In a particular embodiment, prior treatment includes treatment with a drug that is elotuzumab. In a particular embodiment, prior treatment includes treatment with a drug that is panobinostat. In certain embodiments, prior treatment comprises treatment with at least one drug, the at least one of a proteasome inhibitor (PI), an IMiD, and / or an anti-CD38 antibody.In certain embodiments, the prior treatment line comprises treatment with at least one drug, the at least one drug comprising at least one of a PI, an IMiD, and / or an alkylating agent. In certain embodiments, the subject relapsed after the previous treatment line.

[0182] In certain embodiments, multiple myeloma is resistant to one or more of the following: bortezomib, carfilzomib, ixazomib, lenalidomide, pomalidomide, thalidomide, dexamethasone, prednisone, alkylating agents, daratumumab, isatuximab, TAK-079, elotuzumab, and / or panobinostat. In certain embodiments, multiple myeloma is resistant to at least one drug after one or more prior treatment lines. In certain embodiments, the at least one drug to which multiple myeloma is resistant includes IMiD. In some embodiments, IMiD includes lenalidomide, pomalidomide, or thalidomide. In some embodiments, IMiD includes lenalidomide. In preferred embodiments, multiple myeloma is lenalidomide-resistant multiple myeloma. In some embodiments, the IMiD is lenalidomide. In certain embodiments, multiple myeloma is resistant to at least two drugs after prior treatment. In certain embodiments, the at least two drugs to which multiple myeloma is resistant include a proteasome inhibitor (PI) and an IMiD (e.g., lenalidomide). In certain embodiments, multiple myeloma is resistant to at least three drugs after prior treatment lines. In certain embodiments, multiple myeloma is resistant to at least four drugs after prior treatment lines. In certain embodiments, at least four prior treatment lines include treatment with at least one drug, the at least one drug comprising at least one of a PI, an IMiD, an anti-CD38 antibody, and / or an alkylating agent. In certain embodiments, multiple myeloma is resistant to at least five drugs after prior treatment lines.

[0183] In some embodiments, the subject has approximately 10% to 30% myeloid plasma cells before the administration of the CAR-T cells.

[0184] In certain embodiments, bone marrow aspiration or biopsy may be performed for clinical evaluation, or for biomarker evaluation. In certain embodiments, clinical staging (morphology, cytogenetics, and immunohistochemistry or immunofluorescence or flow cytometry) may be performed. In certain embodiments, immunophenotyping may be performed on a portion of the bone marrow aspiration to monitor BCMA, checkpoint ligand expression in CD138-positive multiple myeloma cells, and checkpoint expression on T cells. In certain embodiments, minimal residual disease (MRD) may be monitored in subjects using next-generation sequencing (NGS) of bone marrow aspirate DNA. NGS of bone marrow aspirate DNA is known to those skilled in the art. In certain embodiments, NGS is performed by clonoSeq. In certain embodiments, myeloma clones may be defined using baseline bone marrow aspirate, and MRD negativity may be assessed using post-treatment samples. In certain embodiments, MRD negativity can be determined based on an evaluable sample. In certain embodiments, an evaluable sample is one or more of the following: calibration, quality control, and having sufficient cells to be evaluated at a specific sensitivity level. In some embodiments, the sensitivity level is 10 -6 In a particular embodiment, the sensitivity level is 10 -6 The sensitivity level is 10 -5 In a particular embodiment, the sensitivity level is 10 -4 In a particular embodiment, the sensitivity level is 10 -3 That is the case.

[0185] In certain embodiments, the efficacy of a treatment method for a subject is determined using efficacy criteria based on the International Myeloma Working Group (IMWG), which is summarized in Table 6. In certain embodiments, the efficacy may be classified as stringent complete remission (sCR). In certain embodiments, the efficacy may be classified as complete remission (CR) that is worse than stringent complete remission (sCR). In certain embodiments, the efficacy may be classified as very good partial remission (VGPR) that is worse than complete remission (CR). In certain embodiments, the efficacy may be classified as partial remission (PR) that is worse than very good partial remission (VGPR). In certain embodiments, the efficacy may be classified as minimal response (MR) that is worse than partial remission (PR). In certain embodiments, the efficacy may be classified as stable disease (SD) that is worse than minimal response (MR). In certain embodiments, the efficacy may be classified as progressive disease (PD) that is worse than stable disease.

[0186] In certain embodiments, the tests used for the determination of efficacy criteria based on the International Myeloma Working Group (IMWG) are measurement of myeloma protein (M protein) in serum and urine, serum calcium after albumin correction, bone marrow examination, skeletal examination, and record review of extramedullary plasmacytoma.

[0187] Non-limiting examples of tests for M protein measurement in blood and urine are known to those skilled in the art and include serum quantitative Ig, serum protein electrophoresis (SPEP), serum immunofixation electrophoresis, serum FLC assay, quantification of urinary M protein by electrophoresis in 24-hour urine (UPEP), urinary immunofixation electrophoresis, and serum β2-microglobulin.

[0188] It is known to those skilled in the art that, in order to detect hypercalcemia, serum calcium levels in blood samples should be calculated after correcting for albumin. Although we do not wish to be constrained by theory, calcium binds to albumin, and only unbound (free) calcium is biologically active; therefore, serum calcium levels must be adjusted for abnormal albumin levels ("corrected serum calcium").

[0189] In certain embodiments, a skeletal examination of the skull, the entire spine, the pelvis, the chest, the humerus, the femur, and any one or all of any other bones may be performed and evaluated by either radiographic imaging ("X-rays") or low-dose computed tomography (CT) diagnostic quality scans without intravenous contrast agents, both of which are known to those skilled in the art. In certain embodiments, X-ray or CT scans may be performed locally after T-cell administration and before confirmation of disease progression, whenever clinically indicated based on symptoms, to record response or progression. In certain embodiments, magnetic resonance imaging (MRI) may be used to evaluate bone disease, but it is not a substitute for a skeletal examination. MRI is known to those skilled in the art. In certain embodiments, if a radionuclide bone scan is used in addition to a whole skeletal examination at the time of screening, both methods may be used to record the disease state. Radionuclide bone scans are known to those skilled in the art. In certain embodiments, a radionuclide bone scan and a whole skeletal examination may be performed at the same time. In certain embodiments, radioactive bone scans are not a substitute for whole-skeletal examinations. In certain embodiments, if disease progression is observed in a subject as a symptom of pain caused by bone changes, the disease progression may be recorded by skeletal examination or other X-ray images, depending on the symptoms observed in the subject.

[0190] In certain embodiments, an extramedullary plasmacytoma may be documented by clinical examination or MRI. In certain embodiments, if the use of intravenous contrast agent is not contraindicated, an extramedullary plasmacytoma may be documented by CT scan. In certain embodiments, an extramedullary plasmacytoma may be documented by fusion of positron emission tomography (PET) and CT scan when the CT elements are of sufficient diagnostic quality. In certain embodiments, determination of measurable extramedullary disease sites may be performed, measured, or evaluated locally every 4 weeks for a subject until a confirmed CR or confirmed disease progression occurs. In certain embodiments, evaluation of extramedullary plasmacytoma may be performed every 12 weeks.

[0191] In certain embodiments, for qualifying VGPR or PR or MR, the sum of products of orthogonal diameters of existing extramedullary plasmacytomas may each have decreased by 90% or at least more than 50%. In certain embodiments, for qualifying disease progression, the sum of products of orthogonal diameters of existing extramedullary plasmacytomas must have increased by at least 50%, or the longest diameter of a previous lesion greater than 1 cm on the short axis must have increased by at least 50%, or a new plasmacytoma must have occurred, either. In certain embodiments, for qualifying disease progression when not all existing extramedullary plasmacytomas have been reported, the sum of products of orthogonal diameters of the reported plasmacytomas has increased by at least 50%. In certain embodiments, when study treatment interferes with immunofixation assay, CR may be defined as disappearance of the original M protein associated with multiple myeloma undergoing immunofixation.

[0192] In certain embodiments, the response of a subject to a treatment method is determined in terms of a change in disease burden or tumor burden. Disease burden or tumor burden represents the type of measurable disease in the subject. In some embodiments, the change in tumor burden may be determined in terms of a change in paraprotein levels associated with treatment. In some embodiments, paraprotein is M protein in serum. In some embodiments, paraprotein is M protein in serum. In some embodiments, the change in tumor burden is determined in terms of the difference (dFLC) between free light chains from tumors and free light chains from non-tumors. In some embodiments, the change in tumor burden is determined in terms of the maximum decrease in paraprotein from baseline, i.e., from before CAR-T cell administration. In some embodiments, the change in tumor burden is determined at a median follow-up period of 28 days or more after CAR-T cell administration. In some embodiments, the change in tumor burden is determined at a median follow-up period of 1 month or more after CAR-T cell administration. In some embodiments, the change in tumor burden is determined at a median follow-up period of 3 months or more after CAR-T cell administration. In some embodiments, changes in tumor burden are assessed at a median follow-up period of 6 months or more after CAR-T cell administration. In some embodiments, changes in tumor burden are assessed at a median follow-up period of 9 months or more after CAR-T cell administration. In some embodiments, changes in tumor burden are assessed at a median follow-up period of 12 months or more after CAR-T cell administration.

[0193] In certain embodiments, the subject is re-treated by a second intravenous infusion of a second dose of CAR-T cells. In certain embodiments, the re-treatment dose is 1.0 × 10⁶ per kilogram of the subject's body weight. 5 ~5.0×10 6 Contains 10 CAR-T cells. In a particular embodiment, the retreatment dose is approximately 0.75 × 10¹⁶ per kilogram of the subject's body weight. 5Contains 1 CAR-T cell. In certain embodiments, the subject is retreated when disease progression occurs after the best response, above minimum response, is achieved following the initial infusion of CAR-T cells. In certain embodiments, the time between the initial infusion of CAR-T cells and the detection of disease progression includes at least 6 months.

[0194] In one embodiment, a method is provided for treating a subject having multiple myeloma, comprising administering a composition comprising a therapeutically effective number of chimeric antigen receptor (CAR)-containing T cells to deliver a certain dose of CAR-expressing T cells (CAR-T cells) to the subject by a single intravenous infusion.

[0195] In some embodiments, the subjects received prior treatment with at least one to three prior treatment lines. In some embodiments, the prior treatment line included treatment with at least one drug, the at least one of a proteasome inhibitor (PI), an IMiD, and an anti-CD38 antibody. In some embodiments, the subjects relapsed after the previous treatment line.

[0196] In some embodiments, multiple myeloma is resistant to at least two drugs after the previous line of treatment. In some embodiments, the at least two drugs to which the subject is resistant include proteasome inhibitors (PIs) and IMiDs (e.g., lenalidomide). In some embodiments, multiple myeloma is resistant to at least three drugs after the previous line of treatment. In some embodiments, multiple myeloma is resistant to at least four drugs after the previous line of treatment. In some embodiments, multiple myeloma is resistant to at least five drugs after the previous line of treatment.

[0197] In some embodiments, the subject is over 65 years of age. In some embodiments, the subject is Black or African American. In some embodiments, the subject has received 1 to 3 prior lines of treatment. In some embodiments, the subject has received at least 1 prior line of treatment. In some embodiments, the subject has received at least 2 prior lines of treatment. In some embodiments, the subject has received at least 3 prior lines of treatment. In some embodiments, the subject has received at least 4 prior lines of treatment. In some embodiments, the subject has multiple myeloma or the subject is resistant to 3 classes of drugs, i.e., the multiple myeloma or the subject is triple-class resistant. In some embodiments, the multiple myeloma or the subject is resistant to 5 drugs or pharmaceuticals, i.e., the multiple myeloma or the subject is 5-drug resistant. In some embodiments, the subject has high-risk disease factors, including high-risk cytogenetic abnormalities, soft tissue plasmacytoma, triple-class resistance, or other high-risk disease factors. In some embodiments, the subject has standard-risk cytogenetics. In some embodiments, the subject has high-risk cytogenetics. In some embodiments, the cytogenetic abnormality is a standard-risk cytogenetic abnormality. In some embodiments, the cytogenetic abnormality is a high-risk cytogenetic abnormality. In some embodiments, the subject has one or more high-risk cytogenetic abnormalities selected from the group including Gain / amp(1q), del(17p), t(4;14), t(14;16), or any combination thereof. In some embodiments, the cytogenetic abnormality includes Gain / amp(1q). In some embodiments, the cytogenetic abnormality includes del(17p). In some embodiments, the cytogenetic abnormality includes t(4;14). In some embodiments, the cytogenetic abnormality includes t(14;16). In some embodiments, the subject has two, three, four, five, or more cytogenetic abnormalities. In some embodiments, the subject has at least two cytogenetic abnormalities. In some embodiments, the subject has at least three cytogenetic abnormalities.In some embodiments, the subject has at least four cytogenetic abnormalities. In some embodiments, the subject has at least five cytogenetic abnormalities. In some embodiments, the subject has at least six cytogenetic abnormalities. In some embodiments, the subject has at least seven cytogenetic abnormalities. In some embodiments, the subject or multiple myeloma is characterized as stage III according to the International Staging Classification. In some embodiments, the subject has a soft tissue plasmacytoma. In some embodiments, the subject has about 10% to about 30% myeloid plasma cells prior to the administration of the CAR-T cells. In some embodiments, the subject has about 31% to about 59% myeloid plasma cells prior to the administration of the CAR-T cells. In some embodiments, the subject has about 60% to about 100% myeloid plasma cells prior to the administration of the CAR-T cells. In some embodiments, the subject's BCMA expression in the tumor is below the median of the multiple myeloma patient population or any randomly selected population. In some embodiments, the subjects have BCMA expression in the tumor that is equal to or greater than the median of the multiple myeloma patient population or any randomly selected population. In some embodiments, the subjects have plasmacytoma. In some embodiments, the plasmacytoma is ossicular. In some embodiments, the plasmacytoma is extramedullary. In some embodiments, the plasmacytoma is both ossicular and extramedullary.

[0198] In certain embodiments, this treatment method is effective in achieving a reduction in tumor burden in the subject. In certain embodiments, this treatment method is effective in achieving a reduction in tumor burden of approximately 1% to 100%, 60% to 100%, 65% to 100%, 70% to 100%, 75% to 100%, 80% to 100%, 85% to 100%, 90% to 100%, 92% to 100%, 95% to 100%, 96% to 100%, 97% to 100%, 98% to 100%, or 99% to 100% in the subject. In certain embodiments, this treatment method is effective in achieving a reduction in tumor burden of approximately 100% in the subject. In certain embodiments, this treatment method is effective in achieving a reduction in tumor burden of approximately 1% to 100% in subjects at a rate of approximately 1% to 100%. In certain embodiments, this treatment method is effective in achieving a reduction in tumor burden of approximately 60% to 100% in subjects at a rate of approximately 1% to 100%. In certain embodiments, this treatment method is effective in achieving a reduction in tumor burden of approximately 65% ​​to 100% in subjects at a rate of approximately 1% to 92%. In certain embodiments, this treatment method is effective in achieving a reduction in tumor burden of approximately 70% to 100% in subjects at a rate of approximately 1% to 88%. In certain embodiments, this treatment method is effective in achieving a reduction in tumor burden of approximately 90% to 100% in subjects at a rate of approximately 1% to 88%. In certain embodiments, this treatment method is effective in achieving a reduction of approximately 95% to 100% of tumor burden in subjects at a rate of approximately 1% to 88%. In certain embodiments, this treatment method is effective in achieving a reduction of approximately 99% to 100% of tumor burden in subjects at a rate of approximately 1% to 88%. In certain embodiments, this treatment method is effective in achieving a reduction of approximately 100% of tumor burden in subjects at a rate of approximately 1% to 83%.

[0199] In certain embodiments, this treatment method is effective in achieving or maintaining a minimal residual disease (MRD) negative state in the subject. In certain embodiments, this treatment method is effective in achieving a minimal residual disease (MRD) negative state in the subject. -6It is effective in achieving a minimal residual disease (MRD) negative state at the sensitivity level. In certain embodiments, this treatment method is effective in the subject at 10 -5 It is effective in achieving a minimal residual disease (MRD) negative state at the sensitivity level. In certain embodiments, this treatment method is effective in the subject at 10 -4 It is effective in achieving a minimal residual disease (MRD) negative state at the sensitivity level. In certain embodiments, this treatment method is effective in the subject at 10 -3 The method is effective in achieving minimal residual disease (MRD) negativity at a certain sensitivity level. In certain embodiments, the treatment method is effective in achieving MRD negativity when assessed in bone marrow. In certain embodiments, the treatment method is effective in maintaining MRD negativity when assessed using an evaluable bone marrow sample. In certain embodiments, the treatment method is effective in achieving MRD negativity when assessed using bone marrow DNA. In some embodiments, the method is effective in achieving minimal residual disease (MRD) negativity in the subject as assessed in bone marrow during a follow-up period approximately 28 days or more after the administration of the CAR-T cells, approximately 2 months or more after the administration of the CAR-T cells, approximately 3 months or more after the administration of the CAR-T cells, approximately 6 months or more after the administration of the CAR-T cells, approximately 9 months or more after the administration of the CAR-T cells, or approximately 12 months or more after the administration of the CAR-T cells. In some embodiments, the minimal residual disease (MRD) negative state is achieved during the initial follow-up period, approximately 28 to 179 days after the injection of the CAR-T cells.

[0200] In certain embodiments, this treatment method is effective in maintaining the minimal residual disease (MRD) negative state that was first achieved in the subject. In certain embodiments, this treatment method is effective in maintaining the minimal residual disease (MRD) negative state. -5 It is effective in maintaining an MRD-negative state at the sensitivity level. In a particular embodiment, this treatment method is effective in the subject at 10 -6 It is effective in achieving a minimal residual disease (MRD) negative state at the sensitivity level. In certain embodiments, this treatment method is 10-4 It is effective in maintaining an MRD-negative state at the sensitivity level. In a particular embodiment, this treatment method is 10 -3 It is effective in maintaining an MRD-negative state at a sensitivity level. In certain embodiments, the treatment method is effective in maintaining an MRD-negative state when determined using a bone marrow sample. In certain embodiments, the treatment method is effective in maintaining an MRD-negative state when determined using an evaluable bone marrow sample. In certain embodiments, the treatment method is effective in maintaining an MRD-negative state when determined using bone marrow DNA. In some embodiments, the method is effective in maintaining the minimal residual disease (MRD)-negative state of the subject as determined in the bone marrow during a second follow-up period approximately 29 to 359 days after the administration of the CAR-T cells, approximately 29 to 9 months after the administration of the CAR-T cells, approximately 29 to 6 months after the administration of the CAR-T cells, approximately 29 to 3 months after the administration of the CAR-T cells, or approximately 29 to 2 months after the administration of the CAR-T cells. In some embodiments, the method is effective in maintaining the minimal residual disease (MRD) negative status of the subject as determined in the bone marrow during a second follow-up period approximately 180 to 359 days after the injection of the CAR-T cells. In some embodiments, the method is effective in maintaining the minimal residual disease (MRD) negative status of the subject as determined in the bone marrow during a second follow-up period approximately 360 to 539 days after the injection of the CAR-T cells.

[0201] In certain embodiments, the effectiveness of the treatment method is determined by evaluating the proportion of subjects in an MRD-negative state. In certain embodiments, the effectiveness of the treatment method is determined by 10 -6 The effectiveness of the treatment method is determined by evaluating the proportion of subjects in an MRD-negative state at a sensitivity level of 10. -5 The effectiveness of the treatment method is determined by evaluating the proportion of subjects in an MRD-negative state at a sensitivity level of 10. -4The effectiveness of the treatment method is determined by evaluating the proportion of subjects in an MRD-negative state at a sensitivity level of 10. -3 The effectiveness of the treatment method is determined by evaluating the proportion of subjects who are MRD-negative at the median follow-up period from CAR-T cell administration to approximately 359 days post-CAR-T cell administration, from CAR-T cell administration to approximately 9 months post-CAR-T cell administration, from CAR-T cell administration to approximately 6 months post-CAR-T cell administration, from CAR-T cell administration to approximately 3 months post-CAR-T cell administration, from CAR-T cell administration to approximately 2 months post-CAR-T cell administration, or from CAR-T cell administration to approximately 29 days post-CAR-T cell administration. In some embodiments, the method is performed during the follow-up period approximately 12 months after the injection of the CAR-T cells. -5 A ratio of approximately 44% or less at the sensitivity threshold level, and 10 during the follow-up period approximately 12 months after the administration of the CAR-T cells. -5 At the sensitivity threshold level, approximately 55% of cases occurred during the follow-up period approximately 12 months after the injection of the CAR-T cells. -5 A ratio of approximately 65% ​​or less at the sensitivity threshold level, and 10 during the follow-up period approximately 18 months after the injection of the CAR-T cells. -4 A ratio of approximately 57% or less at the sensitivity threshold level, and 10 during the follow-up period approximately 18 months after the injection of the CAR-T cells. -4 At the sensitivity threshold level, approximately 67% of cases occurred during the follow-up period approximately 18 months after the injection of the CAR-T cells. -4 A ratio of approximately 76% or less at the sensitivity threshold level, and 10 during the follow-up period approximately 18 months after the injection of the CAR-T cells. -5 A ratio of approximately 47% or less at the sensitivity threshold level, and 10 during the follow-up period approximately 18 months after the injection of the CAR-T cells. -5 At the sensitivity threshold level, approximately 58% of cases occurred during the follow-up period approximately 18 months after the injection of the CAR-T cells. -5A ratio of approximately 68% or less at the sensitivity threshold level, and 10 during the follow-up period approximately 18 months after the injection of the CAR-T cells. -6 A ratio of approximately 29% or less at the sensitivity threshold level, and 10 during the follow-up period approximately 18 months after the injection of the CAR-T cells. -6 At the sensitivity threshold level, approximately 39% of cases occurred, or 10% occurred during the follow-up period approximately 18 months after the injection of the CAR-T cells. -6 This method is effective in achieving the minimal residual disease (MRD) negative state at a rate of approximately 50% or less at the sensitivity threshold level. In some embodiments, the method involves a follow-up period of approximately 12 months after the injection of the CAR-T cells. -5 At the sensitivity threshold level, the ratio was approximately 44% to approximately 65%, and during the follow-up period approximately 18 months after the injection of the CAR-T cells, 10 -4 At the sensitivity threshold level, the ratio was approximately 57% to 76%, and during the follow-up period approximately 18 months after the injection of the CAR-T cells, 10 -5 At the sensitivity threshold level, the ratio was approximately 47% to approximately 68%, or 10 during the follow-up period approximately 18 months after the injection of the CAR-T cells. -6 At the sensitivity threshold level, the method is effective in achieving the minimal residual disease (MRD) negative state in a ratio of approximately 29% to approximately 50%. In some embodiments, the method involves administering CAR-T cells during a follow-up period of approximately 12 months after the administration of the CAR-T cells. -5 At the sensitivity threshold level, approximately 55% of cases occurred during the follow-up period approximately 18 months after the injection of the CAR-T cells. -4 At the sensitivity threshold level, approximately 67% of cases occurred during the follow-up period approximately 18 months after the injection of the CAR-T cells. -5 At the sensitivity threshold level, approximately 58% of cases occurred, or 10 cases occurred during the follow-up period approximately 18 months after the injection of the CAR-T cells. -6 At the sensitivity threshold level, it is effective in achieving the minimal residual disease (MRD) negative state in approximately 39% of cases.

[0202] In certain embodiments, the effectiveness of the treatment method is determined by evaluating the proportion of MRD-negative subjects with evaluable bone marrow. In certain embodiments, the effectiveness of the treatment method is determined by 10 -6 It is determined by evaluating the proportion of MRD-negative subjects with bone marrow that can be evaluated at a sensitivity level. In a particular embodiment, the effectiveness of the treatment method is 10 -5 It is determined by evaluating the proportion of MRD-negative subjects with bone marrow that can be evaluated at a sensitivity level. In a particular embodiment, the effectiveness of the treatment method is 10 -4 It is determined by evaluating the proportion of MRD-negative subjects with bone marrow that can be evaluated at a sensitivity level. In a particular embodiment, the effectiveness of the treatment method is 10 -3 The effectiveness of the treatment method is determined by evaluating the proportion of MRD-negative subjects with bone marrow that can be evaluated at a sensitivity level. In certain embodiments, the effectiveness of the treatment method is determined by evaluating the proportion of MRD-negative subjects with bone marrow that can be evaluated at a median follow-up period during the following periods: from CAR-T cell administration to approximately 359 days post-CAR-T cell administration, from CAR-T cell administration to approximately 9 months post-CAR-T cell administration, from CAR-T cell administration to approximately 6 months post-CAR-T cell administration, from CAR-T cell administration to approximately 3 months post-CAR-T cell administration, from CAR-T cell administration to approximately 2 months post-CAR-T cell administration, or from CAR-T cell administration to approximately 29 days post-CAR-T cell administration. In some embodiments, the method is performed during a follow-up period approximately 12 months after the infusion of the CAR-T cells. -5 The proportion of subjects with a sample that can be evaluated at the sensitivity threshold level is approximately 83% or less, and 10% of the subjects during the follow-up period approximately 12 months after the injection of the CAR-T cells. -5 Approximately 93% of subjects with samples that can be evaluated at the sensitivity threshold level, and 10 during the follow-up period approximately 12 months after the injection of the CAR-T cells. -5 The ratio of approximately 98% or less of subjects with a sample that can be evaluated at the sensitivity threshold level, and 10 during the follow-up period approximately 18 months after the injection of the CAR-T cells. -5The proportion of subjects with a sample that can be evaluated at the sensitivity threshold level is approximately 82% or less, and 10% of the subjects during the follow-up period approximately 18 months after the injection of the CAR-T cells. -5 Approximately 92% of subjects with samples that can be evaluated at the sensitivity threshold level, or 10 during the follow-up period approximately 18 months after the injection of the CAR-T cells. -5 This method is effective in achieving minimal residual disease (MRD) negativity in approximately 97% or less of subjects with samples that can be evaluated at the sensitivity threshold level. In some embodiments, the method is performed during a follow-up period of approximately 12 months after the injection of the CAR-T cells. -5 Approximately 83% to 98% of subjects with samples that can be evaluated at the sensitivity threshold level, or 10 during the follow-up period approximately 18 months after the injection of the CAR-T cells. -5 The method is effective in achieving minimal residual disease (MRD) negativity in approximately 82% to 97% of subjects with samples that can be evaluated at the sensitivity threshold level. In some embodiments, the method is performed during a follow-up period of approximately 12 months after the injection of the CAR-T cells. -5 Approximately 93% of subjects with samples that can be evaluated at the sensitivity threshold level, or 10 during the follow-up period approximately 18 months after the injection of the CAR-T cells. -5 This method is effective in achieving minimal residual disease (MRD) negativity in approximately 92% of subjects with samples that can be evaluated at the sensitivity threshold level.

[0203] In some embodiments, the method is effective in achieving at least one response in a subject after the injection of the CAR-T cells, the at least one response including, in order from best to worst, a severe complete response, a complete response, a best partial response, a partial response, or a minimal response.

[0204] In some embodiments, the method is effective in achieving an initial response within approximately 27 days, 29 days, 42 days, 89 days, or 321 days or more after the injection of the CAR-T cells. In some embodiments, the method is effective in achieving an initial response between approximately 27 and 321 days after the injection of the CAR-T cells. In some embodiments, the method is effective in achieving an initial response between approximately 27 and 89 days after the injection of the CAR-T cells. In some embodiments, the method is effective in achieving an initial response within approximately 42 days after the injection of the CAR-T cells. In some embodiments, the method is effective in achieving an initial response within approximately 29 days after the injection of the CAR-T cells.

[0205] In certain embodiments, the effectiveness of the treatment method is determined by evaluating the percentage of subjects who achieve a strict complete response. In certain embodiments, the effectiveness of the treatment method is determined by evaluating the percentage of subjects who achieve a complete response or better. In certain embodiments, the effectiveness of the treatment method is determined by evaluating the percentage of subjects who achieve a best partial response or better. In certain embodiments, the effectiveness of the treatment method is determined by evaluating the percentage of subjects who achieve a partial response or better. In certain embodiments, the effectiveness of the treatment method is determined by evaluating the percentage of subjects who achieve a minimum response or better.

[0206] In some embodiments, the method is effective in achieving one of the best responses, i.e., a best response greater than or equal to a minimum response, among minimal response, partial response, best partial response, complete response, or strict complete response. In some embodiments, the percentage in which the method is effective in achieving a best response greater than or equal to a minimum response is called the clinical benefit rate. In some embodiments, the method is effective in achieving a best response greater than or equal to a minimum response in a ratio of about 91% or less during a follow-up period about 12 months after the injection of the CAR-T cells, a ratio of about 97% or less during a follow-up period about 12 months after the injection of the CAR-T cells, a ratio of about 99% or less during a follow-up period about 12 months after the injection of the CAR-T cells, a ratio of about 93% or less during a follow-up period about 18 months after the injection of the CAR-T cells, a ratio of about 98% or less during a follow-up period about 18 months after the injection of the CAR-T cells, or a ratio of about 100% or less during a follow-up period about 18 months after the injection of the CAR-T cells. In some embodiments, the method is effective in achieving one of the best responses—minimal response, partial response, best partial response, complete response, or severe complete response—at a rate of approximately 91% to 99% during a follow-up period approximately 12 months after the injection of the CAR-T cells, or at a rate of approximately 93% to 100% during a follow-up period approximately 18 months after the injection of the CAR-T cells. In some embodiments, the method is effective in achieving one of the best responses—minimal response, partial response, best partial response, complete response, or severe complete response—at a rate of approximately 97% during a follow-up period approximately 12 months after the injection of the CAR-T cells, or at a rate of approximately 98% during a follow-up period approximately 18 months after the injection of the CAR-T cells.

[0207] In some embodiments, this method is effective in achieving strict complete remission at a rate of approximately 40% to approximately 90%. In some embodiments, this method is effective in achieving strict complete remission at a rate of approximately 50% to approximately 80%. In some embodiments, this method is effective in achieving strict complete remission at a rate of approximately 58.2%. In some embodiments, this method is effective in achieving strict complete remission at a rate of approximately 68.8%.

[0208] In some embodiments, this method is effective in achieving complete remission in a rate of approximately 10% to approximately 20%. In some embodiments, this method is effective in achieving complete remission in a rate of approximately 14.9%. In some embodiments, this method is effective in achieving complete remission in a rate of approximately 17.6%.

[0209] In some embodiments, this method is effective in achieving the best partial response.

[0210] In some embodiments, this method is effective in achieving a partial response.

[0211] In some embodiments, the method is effective in achieving the best efficacy, i.e., an efficacy better than a partial efficacy, which is any one of a partial efficacy, a best partial efficacy, a complete efficacy, or a strict complete efficacy. In some embodiments, the ratio at which the method is effective in achieving an efficacy better than a partial efficacy is referred to as an overall survival rate or an overall efficacy rate. In some embodiments, the method is effective in achieving an efficacy better than a partial efficacy at a ratio of about 91% or less, at a ratio of about 97% or less, or at a ratio of about 99% or less during a follow-up period about 12 months after the injection of the CAR-T cells; at a ratio of about 93% or less, at a ratio of about 97% or less, or at a ratio of about 100% or less during a follow-up period about 18 months after the injection of the CAR-T cells. In some embodiments, the method is effective in achieving the best efficacy, which is any one of a partial efficacy, a best partial efficacy, a complete efficacy, or a strict complete efficacy, at a ratio of about 91% to about 99% during a follow-up period about 12 months after the injection of the CAR-T cells or at a ratio of about 93% to about 100% during a follow-up period about 18 months after the injection of the CAR-T cells. In some embodiments, the method is effective in achieving the best efficacy, which is any one of a partial efficacy, a best partial efficacy, a complete efficacy, or a strict complete efficacy, at a ratio of about 97% during a follow-up period about 12 months after the injection of the CAR-T cells or at a ratio of about 97% during a follow-up period about 18 months after the injection of the CAR-T cells.

[0212] In some embodiments, the method is effective in achieving one of the best responses, which is either the best partial response, the complete response, or the severe complete response, i.e., a best response of the best partial response or better. In some embodiments, the method is effective in achieving a best response of the best partial response or better in a ratio of about 86% or less during a follow-up period about 12 months after the injection of the CAR-T cells, a ratio of about 93% or less during a follow-up period about 12 months after the injection of the CAR-T cells, a ratio of about 97% or less during a follow-up period about 12 months after the injection of the CAR-T cells, a ratio of about 88% or less during a follow-up period about 18 months after the injection of the CAR-T cells, a ratio of about 95% or less during a follow-up period about 18 months after the injection of the CAR-T cells, or a ratio of about 98% or less during a follow-up period about 18 months after the injection of the CAR-T cells. In some embodiments, the method is effective in achieving one of the best responses, either a best partial response, a complete response, or a severe complete response, at a rate of approximately 86% to 97% during a follow-up period approximately 12 months after the injection of the CAR-T cells, or at a rate of approximately 88% to 98% during a follow-up period approximately 18 months after the injection of the CAR-T cells. In some embodiments, the method is effective in achieving one of the best responses, either a best partial response, a complete response, or a severe complete response, at a rate of approximately 93% during a follow-up period approximately 12 months after the injection of the CAR-T cells, or at a rate of approximately 95% during a follow-up period approximately 18 months after the injection of the CAR-T cells.

[0213] In some embodiments, the method is effective in achieving complete response or the best response of strict complete response, i.e., the best response of complete response or better. In some embodiments, the method is effective in achieving the best response of complete response or better at a rate of approximately 57% or less during the follow-up period approximately 12 months after the injection of the CAR-T cells, approximately 67% or less during the follow-up period approximately 12 months after the injection of the CAR-T cells, approximately 76% or less during the follow-up period approximately 12 months after the injection of the CAR-T cells, approximately 73% or less during the follow-up period approximately 18 months after the injection of the CAR-T cells, approximately 83% or less during the follow-up period approximately 18 months after the injection of the CAR-T cells, or approximately 89% or less during the follow-up period approximately 18 months after the injection of the CAR-T cells. In some embodiments, the method is effective in achieving the best response, which is complete response or severe complete response, at a rate of approximately 57% to approximately 76% during a follow-up period approximately 12 months after the injection of the CAR-T cells, or at a rate of approximately 73% to approximately 89% during a follow-up period approximately 18 months after the injection of the CAR-T cells. In some embodiments, the method is effective in achieving the best response, which is complete response or severe complete response, at a rate of approximately 67% during a follow-up period approximately 12 months after the injection of the CAR-T cells, or at a rate of approximately 83% during a follow-up period approximately 18 months after the injection of the CAR-T cells.

[0214] In some embodiments, the method is effective in achieving the best response of strict complete response. In some embodiments, the method is effective in achieving the best response of strict complete response at a rate of approximately 57% or less at a follow-up period approximately 12 months after the injection of the CAR-T cells, at a rate of approximately 67% or less at a follow-up period approximately 12 months after the injection of the CAR-T cells, at a rate of approximately 76% or less at a follow-up period approximately 12 months after the injection of the CAR-T cells, at a rate of approximately 73% or less at a follow-up period approximately 18 months after the injection of the CAR-T cells, at a rate of approximately 83% or less at a follow-up period approximately 18 months after the injection of the CAR-T cells, or at a rate of approximately 89% or less at a follow-up period approximately 18 months after the injection of the CAR-T cells. In some embodiments, the method is effective in achieving the best response of strict complete response at a rate of approximately 57% to approximately 76% during a follow-up period approximately 12 months after the injection of the CAR-T cells, or at a rate of approximately 73% to approximately 89% during a follow-up period approximately 18 months after the injection of the CAR-T cells. In some embodiments, the method is effective in achieving the best response of strict complete response at a rate of approximately 67% during a follow-up period approximately 12 months after the injection of the CAR-T cells, or at a rate of approximately 83% during a follow-up period approximately 18 months after the injection of the CAR-T cells.

[0215] In some embodiments, the method is effective in achieving the best response within approximately 27 days, 78 days, 153 days, 293 days, or 534 days or more after the injection of the CAR-T cells. In some embodiments, the method is effective in achieving the best response between approximately 27 and 534 days after the injection of the CAR-T cells. In some embodiments, the method is effective in achieving the best response between approximately 27 and 293 days after the injection of the CAR-T cells. In some embodiments, the method is effective in achieving the best response within approximately 153 days after the injection of the CAR-T cells. In some embodiments, the method is effective in achieving the best response within approximately 78 days after the injection of the CAR-T cells.

[0216] In some embodiments, the method is effective in maintaining the response during follow-up periods from the time of the initial response to approximately 180 days after the injection of the CAR-T cells, from the time of the initial response to approximately 357 days after the injection of the CAR-T cells, from the time of the initial response to approximately 606 days after the injection of the CAR-T cells, or from the time of the initial response to approximately 654 days after the injection of the CAR-T cells. In some embodiments, the method involves a ratio of approximately 77% or less during the follow-up period approximately 6 months after the injection of the CAR-T cells, a ratio of approximately 85% or less during the follow-up period approximately 6 months after the injection of the CAR-T cells, a ratio of approximately 91% or less during the follow-up period approximately 6 months after the injection of the CAR-T cells, a ratio of approximately 63% or less during the follow-up period approximately 12 months after the injection of the CAR-T cells, a ratio of approximately 74% or less during the follow-up period approximately 12 months after the injection of the CAR-T cells, a ratio of approximately 81% or less during the follow-up period approximately 12 months after the injection of the CAR-T cells, a ratio of approximately 56% or less during the follow-up period approximately 18 months after the injection of the CAR-T cells, and approximately 18 months after the injection of the CAR-T cells It is effective in maintaining efficacy at a rate of approximately 67% or less during the follow-up period, approximately 75% or less during the follow-up period approximately 18 months after the injection of the CAR-T cells, approximately 52% or less during the follow-up period approximately 21 months after the injection of the CAR-T cells, approximately 63% or less during the follow-up period approximately 21 months after the injection of the CAR-T cells, approximately 72% or less during the follow-up period approximately 21 months after the injection of the CAR-T cells, approximately 48% or less during the follow-up period approximately 24 months after the injection of the CAR-T cells, approximately 60% or less during the follow-up period approximately 24 months after the injection of the CAR-T cells, or approximately 70% or less during the follow-up period approximately 24 months after the injection of the CAR-T cells.In some embodiments, the method is effective in maintaining efficacy at a rate of approximately 77% to 91% during a follow-up period approximately 6 months after the injection of CAR-T cells, approximately 63% to 81% during a follow-up period approximately 12 months after the injection of CAR-T cells, approximately 56% to 75% during a follow-up period approximately 18 months after the injection of CAR-T cells, approximately 52% to 72% during a follow-up period approximately 21 months after the injection of CAR-T cells, or approximately 48% to 70% during a follow-up period approximately 24 months after the injection of CAR-T cells. In some embodiments, the method is effective in maintaining efficacy in approximately 85% of cases during a follow-up period approximately 6 months after the injection of CAR-T cells, approximately 74% during a follow-up period approximately 12 months after the injection of CAR-T cells, approximately 67% during a follow-up period approximately 18 months after the injection of CAR-T cells, approximately 63% during a follow-up period approximately 21 months after the injection of CAR-T cells, or approximately 60% during a follow-up period approximately 24 months after the injection of CAR-T cells.

[0217] In some embodiments, the method described herein involves 10 times between the time of administration of the CAR-T cells and approximately 3 months after the administration of the CAR-T cells. -5The method is effective in achieving a minimal residual disease (MRD)-negative state of the subject as determined in the bone marrow at the sensitivity threshold level. In some embodiments, the method is effective in achieving either a minimal residual disease (MRD)-negative complete response or a minimal residual disease (MRD)-negative severe complete response in a ratio of approximately 25% or less during a follow-up period approximately 12 months after the injection of the CAR-T cells, approximately 34% or less during a follow-up period approximately 12 months after the injection of the CAR-T cells, approximately 44% or less during a follow-up period approximately 12 months after the injection of the CAR-T cells, approximately 33% or less during a follow-up period approximately 18 months after the injection of the CAR-T cells, approximately 43% or less during a follow-up period approximately 18 months after the injection of the CAR-T cells, or approximately 54% or less during a follow-up period approximately 18 months after the injection of the CAR-T cells. In some embodiments, the method is effective in achieving either a minimal residual disease (MRD)-negative complete response or a minimal residual disease (MRD)-negative severe complete response in about 25% to about 44% of cases during a follow-up period about 12 months after the injection of the CAR-T cells, or in about 33% to about 54% of cases during a follow-up period about 18 months after the injection of the CAR-T cells. In some embodiments, the method is effective in achieving either a minimal residual disease (MRD)-negative complete response or a minimal residual disease (MRD)-negative severe complete response in about 34% of cases during a follow-up period about 12 months after the injection of the CAR-T cells, or in about 43% of cases during a follow-up period about 18 months after the injection of the CAR-T cells.

[0218] In some embodiments, the method is effective in achieving progression-free survival of the subject. In some embodiments, the method is effective in achieving progression-free survival of the subject at a time between the injection of the CAR-T cells and approximately 209 days after the injection of the CAR-T cells, between the injection of the CAR-T cells and approximately 386 days after the injection of the CAR-T cells, between the injection of the CAR-T cells and approximately 632 days after the injection of the CAR-T cells, or between the injection of the CAR-T cells and approximately 684 days after the injection of the CAR-T cells. In some embodiments, the method involves a follow-up period of approximately 6 months after the injection of CAR-T cells with a rate of approximately 79% or more, approximately 88% or more, approximately 93% or more, approximately 12 months after the injection of CAR-T cells with a rate of approximately 67% or more, approximately 76% or more, approximately 84% or more, approximately 18 months after the injection of CAR-T cells with a rate of approximately 57% or more, and approximately 18 months after the injection of CAR-T cells with a rate of approximately 79% or more. It is effective in achieving progression-free survival at a rate of approximately 67% or more during the low-up period, approximately 75% or more during the follow-up period approximately 18 months after the injection of the CAR-T cells, approximately 57% or more during the follow-up period approximately 21 months after the injection of the CAR-T cells, approximately 67% or more during the follow-up period approximately 21 months after the injection of the CAR-T cells, approximately 75% or more during the follow-up period approximately 21 months after the injection of the CAR-T cells, approximately 49% or more during the follow-up period approximately 24 months after the injection of the CAR-T cells, approximately 61% or more during the follow-up period approximately 24 months after the injection of the CAR-T cells, or approximately 70% or more during the follow-up period approximately 24 months after the injection of the CAR-T cells.In some embodiments, the method is effective in achieving progression-free survival at a rate of approximately 79% to 93% during a follow-up period approximately 6 months after the injection of CAR-T cells, approximately 67% to 84% during a follow-up period approximately 12 months after the injection of CAR-T cells, approximately 57% to 75% during a follow-up period approximately 18 months after the injection of CAR-T cells, approximately 57% to 75% during a follow-up period approximately 21 months after the injection of CAR-T cells, or approximately 49% to 70% during a follow-up period approximately 24 months after the injection of CAR-T cells. In some embodiments, the method is effective in achieving progression-free survival at a rate of approximately 88% during a follow-up period approximately 6 months after the injection of CAR-T cells, approximately 76% during a follow-up period approximately 12 months after the injection of CAR-T cells, approximately 67% during a follow-up period approximately 18 months after the injection of CAR-T cells, approximately 67% during a follow-up period approximately 21 months after the injection of CAR-T cells, or approximately 61% during a follow-up period approximately 24 months after the injection of CAR-T cells.

[0219] In some embodiments, the method further includes treating the subject with respect to cytokine release syndrome about one day after the injection of the CAR-T cells. In some embodiments, the method is effective in achieving a recovery rate from cytokine release syndrome of about 1% to about 99% at about 1, 3, 4, 6, 16, or 97 days after the onset of the cytokine release syndrome.

[0220] In some embodiments, the method further includes treating the subject for immunoeffector cell-associated neurotoxicity about three days after the injection of the CAR-T cells. In some embodiments, the method is effective in achieving a recovery rate from immunoeffector cell-associated neurotoxicity of about 1% to about 17% at about 1, 4, 5, 8, 12, or 16 days after the onset of the immunoeffector cell-associated neurotoxicity.

[0221] In certain embodiments, this treatment method is effective in achieving a reduction in tumor burden in subjects. In certain embodiments, this treatment method is effective in achieving a reduction in tumor burden in more than 90% of subjects. In certain embodiments, this treatment method is effective in achieving a reduction in tumor burden in more than 91% of subjects. In certain embodiments, this treatment method is effective in achieving a reduction in tumor burden in more than 92% of subjects. In certain embodiments, this treatment method is effective in achieving a reduction in tumor burden in more than 93% of subjects. In certain embodiments, this treatment method is effective in achieving a reduction in tumor burden in more than 94% of subjects. In certain embodiments, this treatment method is effective in achieving a reduction in tumor burden in more than 95% of subjects. In certain embodiments, this treatment method is effective in achieving a reduction in tumor burden in more than 96% of subjects. In certain embodiments, this treatment method is effective in achieving a reduction in tumor burden in more than 97% of subjects. In certain embodiments, this treatment method is effective in achieving a reduction in tumor burden in more than 98% of subjects. In certain embodiments, this treatment method is effective in achieving a reduction in tumor burden in over 99% of subjects. In some embodiments, this treatment method is effective in achieving a reduction in tumor burden in 100% of subjects.

[0222] In certain embodiments, this treatment method is effective in achieving or maintaining a minimal residual disease (MRD) negative state in the subject. In certain embodiments, this treatment method is effective in achieving a minimal residual disease (MRD) negative state in the subject. In certain embodiments, this treatment method is effective in achieving a minimal residual disease (MRD) negative state in the subject. -6 It is effective in achieving a minimal residual disease (MRD) negative state at the sensitivity level. In certain embodiments, this treatment method is effective in the subject at 10 -5 It is effective in achieving a minimal residual disease (MRD) negative state at the sensitivity level. In certain embodiments, this treatment method is effective in the subject at 10 -4It is effective in achieving a minimal residual disease (MRD) negative state at the sensitivity level. In certain embodiments, this treatment method is effective in the subject at 10 -3 It is effective in achieving minimal residual disease (MRD) negativity at a certain sensitivity level. In certain embodiments, this treatment method is effective in achieving MRD negativity when assessed using bone marrow. In certain embodiments, this treatment method is effective in maintaining MRD negativity when assessed using an evaluable bone marrow sample. In certain embodiments, this treatment method is effective in achieving MRD negativity when assessed using bone marrow DNA. In certain embodiments, this treatment method is effective in achieving MRD negativity when assessed during a follow-up period of 28 days or more after CAR-T cell administration. In certain embodiments, this treatment method is effective in achieving MRD negativity when assessed during a follow-up period of 1 month or more after CAR-T cell administration. In certain embodiments, this treatment method is effective in achieving MRD negativity when assessed during a follow-up period of 3 months or more after CAR-T cell administration. In certain embodiments, this treatment method is effective in achieving MRD negativity when assessed during a follow-up period of 6 months or more after CAR-T cell administration. In certain embodiments, this treatment method is effective in achieving an MRD-negative state when assessed during a follow-up period of 9 months or more after CAR-T cell administration. In certain embodiments, this treatment method is effective in achieving an MRD-negative state when assessed during a follow-up period of 12 months or more after CAR-T cell administration.

[0223] In certain embodiments, this treatment method is effective in maintaining the minimal residual disease (MRD) negative state that was first achieved in the subject. In certain embodiments, this treatment method is effective in maintaining the minimal residual disease (MRD) negative state. -5 It is effective in maintaining an MRD-negative state at the sensitivity level. In a particular embodiment, this treatment method is effective in the subject at 10 -6 It is effective in achieving a minimal residual disease (MRD) negative state at the sensitivity level. In certain embodiments, this treatment method is 10 -4It is effective in maintaining an MRD-negative state at the sensitivity level. In a particular embodiment, this treatment method is 10 -3 It is effective in maintaining an MRD-negative state at a sensitivity level. In certain embodiments, this treatment method is effective in maintaining an MRD-negative state when determined using a bone marrow sample. In certain embodiments, this treatment method is effective in maintaining an MRD-negative state when determined using an evaluable bone marrow sample. In certain embodiments, this treatment method is effective in maintaining an MRD-negative state when determined using bone marrow DNA. In certain embodiments, this treatment method is effective in maintaining an MRD-negative state when determined during a follow-up period of one month or more after CAR-T cell administration. In certain embodiments, this treatment method is effective in maintaining an MRD-negative state when determined during a follow-up period of three months or more after CAR-T cell administration. In certain embodiments, this treatment method is effective in maintaining an MRD-negative state when determined during a follow-up period of six months or more after CAR-T cell administration. In certain embodiments, this treatment method is effective in maintaining an MRD-negative state when determined during a follow-up period of nine months or more after CAR-T cell administration. In certain embodiments, this treatment method is effective in maintaining an MRD-negative state when assessed during a follow-up period of 12 months or more after CAR-T cell administration.

[0224] In certain embodiments, the effectiveness of the treatment method is determined by evaluating the proportion of subjects in an MRD-negative state. In certain embodiments, the effectiveness of the treatment method is determined by 10 -6 The effectiveness of the treatment method is determined by evaluating the proportion of subjects in an MRD-negative state at a sensitivity level of 10. -5 The effectiveness of the treatment method is determined by evaluating the proportion of subjects in an MRD-negative state at a sensitivity level of 10. -4 The effectiveness of the treatment method is determined by evaluating the proportion of subjects in an MRD-negative state at a sensitivity level of 10. -3The effectiveness of the treatment method is determined by evaluating the proportion of subjects who are MRD-negative at a sensitivity level. In certain embodiments, the effectiveness of the treatment method is determined by evaluating the proportion of subjects who are MRD-negative at a median follow-up period of 28 days or more after CAR-T cell administration. In certain embodiments, the effectiveness of the treatment method is determined by evaluating the proportion of subjects who are MRD-negative at a median follow-up period of 1 month or more after CAR-T cell administration. In certain embodiments, the effectiveness of the treatment method is determined by evaluating the proportion of subjects who are MRD-negative at a median follow-up period of 3 months or more after CAR-T cell administration. In certain embodiments, the effectiveness of the treatment method is determined by evaluating the proportion of subjects who are MRD-negative at a median follow-up period of 6 months or more after CAR-T cell administration. In certain embodiments, the effectiveness of the treatment method is determined by evaluating the proportion of subjects who are MRD-negative at a median follow-up period of 9 months or more after CAR-T cell administration. In certain embodiments, the effectiveness of the treatment method is determined by evaluating the proportion of subjects who are MRD-negative at a median follow-up period of 12 months or more after CAR-T cell administration.

[0225] In certain embodiments, the effectiveness of the treatment method is determined by evaluating the proportion of MRD-negative subjects with evaluable bone marrow. In certain embodiments, the effectiveness of the treatment method is determined by 10 -6 It is determined by evaluating the proportion of MRD-negative subjects with bone marrow that can be evaluated at a sensitivity level. In a particular embodiment, the effectiveness of the treatment method is 10 -5 It is determined by evaluating the proportion of MRD-negative subjects with bone marrow that can be evaluated at a sensitivity level. In a particular embodiment, the effectiveness of the treatment method is 10 -4 It is determined by evaluating the proportion of MRD-negative subjects with bone marrow that can be evaluated at a sensitivity level. In a particular embodiment, the effectiveness of the treatment method is 10 -3The effectiveness of the treatment method is determined by evaluating the proportion of MRD-negative subjects with bone marrow that can be evaluated at a sensitivity level. In certain embodiments, the effectiveness of the treatment method is determined by evaluating the proportion of MRD-negative subjects with bone marrow that can be evaluated at a median follow-up period of 28 days or more after CAR-T cell administration. In certain embodiments, the effectiveness of the treatment method is determined by evaluating the proportion of MRD-negative subjects with bone marrow that can be evaluated at a median follow-up period of 1 month or more after CAR-T cell administration. In certain embodiments, the effectiveness of the treatment method is determined by evaluating the proportion of MRD-negative subjects with bone marrow that can be evaluated at a median follow-up period of 3 months or more after CAR-T cell administration. In certain embodiments, the effectiveness of the treatment method is determined by evaluating the proportion of MRD-negative subjects with bone marrow that can be evaluated at a median follow-up period of 6 months or more after CAR-T cell administration. In certain embodiments, the effectiveness of the treatment method is determined by evaluating the proportion of MRD-negative subjects with bone marrow that can be evaluated at a median follow-up period of 9 months or more after CAR-T cell administration. In certain embodiments, the effectiveness of the treatment method is determined by evaluating the proportion of MRD-negative subjects with evaluable bone marrow at a median follow-up period of 12 months or more after CAR-T cell administration.

[0226] In certain embodiments, the effectiveness of a treatment method is determined by evaluating the proportion of subjects with a strict complete response. In certain embodiments, the effectiveness of a treatment method is determined by evaluating the proportion of subjects with a complete response or better. In certain embodiments, the effectiveness of a treatment method is determined by evaluating the proportion of subjects with a best partial response or better. In certain embodiments, the effectiveness of a treatment method is determined by evaluating the proportion of subjects with a partial response or better. In certain embodiments, the effectiveness of a treatment method is determined using the overall response rate. In some embodiments, the overall response rate is the proportion of subjects with a partial response or better.

[0227] In a particular embodiment, the method is 10 -5This method is effective in achieving a minimal residual disease (MRD) negativity rate higher than 39% at the sensitivity threshold level. In certain embodiments, this method is 10 -5 This method is effective in achieving a minimal residual disease (MRD) negativity rate higher than 44% at the sensitivity threshold level. In certain embodiments, this method is 10 -5 This method is effective in achieving a minimal residual disease (MRD) negativity rate higher than 49% at the sensitivity threshold level. In certain embodiments, this method is 10 -5 This method is effective in achieving a minimal residual disease (MRD) negativity rate higher than 54% at the sensitivity threshold level. In certain embodiments, this method is 10 -5 This method is effective in achieving a minimal residual disease (MRD) negativity rate higher than 59% at the sensitivity threshold level. In certain embodiments, this method is 10 -5 This method is effective in achieving a minimal residual disease (MRD) negativity rate higher than 64% at the sensitivity threshold level. In certain embodiments, this method is 10 -5 This method is effective in achieving a minimal residual disease (MRD) negativity rate higher than 69% at the sensitivity threshold level. In certain embodiments, this method is 10 -5 This method is effective in achieving a minimal residual disease (MRD) negativity rate higher than 74% at the sensitivity threshold level. In certain embodiments, this method is 10 -5 This method is effective in achieving a minimal residual disease (MRD) negativity rate higher than 70% at the sensitivity threshold level. In certain embodiments, the method is used to evaluate bone marrow. -5 This method is effective in achieving a minimal residual disease (MRD) negativity rate higher than 75% at the sensitivity threshold level. In certain embodiments, the method is used to evaluate bone marrow. -5 This method is effective in achieving a minimal residual disease (MRD) negativity rate higher than 80% at the sensitivity threshold level. In certain embodiments, the method is used to evaluate bone marrow at 10 -5 This method is effective in achieving a minimal residual disease (MRD) negativity rate higher than 85% at the sensitivity threshold level. In certain embodiments, the method is used to evaluate bone marrow at 10 -5This method is effective in achieving a minimal residual disease (MRD) negativity rate higher than 90% at the sensitivity threshold level. In certain embodiments, the method is used to evaluate bone marrow. -5 This method is effective in achieving a minimal residual disease (MRD) negativity rate higher than 95% at the sensitivity threshold level. In certain embodiments, the method is used to evaluate bone marrow at 10 -5 It is effective in achieving a 100% minimal residual disease (MRD) negativity rate at the sensitivity threshold level.

[0228] In certain embodiments, this treatment method is effective in achieving an overall response rate of more than 75%. In certain embodiments, this treatment method is effective in achieving an overall response rate of more than 80%. In certain embodiments, this treatment method is effective in achieving an overall response rate of more than 85%. In certain embodiments, this treatment method is effective in achieving an overall response rate of more than 90%. In certain embodiments, this treatment method is effective in achieving an overall response rate of more than 91%. In certain embodiments, this method is effective in achieving an overall response rate of more than 93%. In certain embodiments, this method is effective in achieving an overall response rate of more than 95%. In certain embodiments, this method is effective in achieving an overall response rate of more than 97%. In certain embodiments, this method is effective in achieving an overall response rate of more than 99%. In some embodiments, this method is effective in achieving an overall response rate of 100%. In certain embodiments, this treatment method is effective in achieving an overall response rate of approximately 84.6%. In certain embodiments, this treatment method is effective in achieving an overall response rate of approximately 99.4%. In certain embodiments, the overall response rate is determined at a median follow-up period of at least 6 months after the injection of the CAR-T cells. In certain embodiments, the overall response rate is determined at a median follow-up period of at least 12 months after the injection of the CAR-T cells.

[0229] In certain embodiments, this treatment method was effective in over 70% of subjects 9 months after administration of CAR-T cells. In certain embodiments, this treatment method was effective in over 72% of subjects 9 months after administration of CAR-T cells. In certain embodiments, this treatment method was effective in over 74% of subjects 9 months after administration of CAR-T cells. In certain embodiments, this treatment method was effective in over 76% of subjects 9 months after administration of CAR-T cells. In certain embodiments, this treatment method was effective in over 78% of subjects 9 months after administration of CAR-T cells. In certain embodiments, this treatment method was effective in over 80% of subjects 9 months after administration of CAR-T cells. In certain embodiments, this treatment method was effective in over 82% of subjects 9 months after administration of CAR-T cells. In certain embodiments, this treatment method was effective in over 84% of subjects 9 months after administration of CAR-T cells. In certain embodiments, this treatment method was effective in over 86% of subjects 9 months after administration of CAR-T cells.

[0230] In certain embodiments, this treatment method was effective in over 54% of responding patients 12 months after CAR-T cell administration. In certain embodiments, this treatment method was effective in over 58% of responding patients 12 months after CAR-T cell administration. In certain embodiments, this treatment method was effective in over 62% of responding patients 12 months after CAR-T cell administration. In certain embodiments, this treatment method was effective in over 66% of responding patients 12 months after CAR-T cell administration. In certain embodiments, this treatment method was effective in over 70% of responding patients 12 months after CAR-T cell administration. In certain embodiments, this treatment method was effective in over 74% of responding patients 12 months after CAR-T cell administration. In certain embodiments, this treatment method was effective in over 78% of responding patients 12 months after CAR-T cell administration.

[0231] In certain embodiments, this treatment method is effective in achieving response periods exceeding or longer than 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 months. In certain embodiments, this treatment method is effective in achieving response periods exceeding 12.4 months. In certain embodiments, this treatment method is effective in achieving response periods exceeding 15.9 months.

[0232] In certain embodiments, this treatment method is effective in achieving a median duration of response of 9 months, 10 months, 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, or longer. In certain embodiments, this treatment method is effective in achieving a median duration of response of 12.4 months or longer. In certain embodiments, this treatment method is effective in achieving a median duration of response of 15.9 months or longer.

[0233] In certain embodiments, this treatment method is effective in achieving complete response or better in more than 60% of subjects. In certain embodiments, this treatment method is effective in achieving complete response or better in more than 61% of subjects. In certain embodiments, this treatment method is effective in achieving complete response or better in more than 62% of subjects. In certain embodiments, this treatment method is effective in achieving complete response or better in more than 63% of subjects. In certain embodiments, this treatment method is effective in achieving complete response or better in more than 64% of subjects. In certain embodiments, this treatment method is effective in achieving complete response or better in more than 65% of subjects. In certain embodiments, this treatment method is effective in achieving complete response or better in more than 66% of subjects. In certain embodiments, this treatment method is effective in achieving complete response or better in more than 67% of subjects. In certain embodiments, complete response or better is determined less than one month after administration of CAR-T cells. In certain embodiments, complete response or better is determined less than three months after administration of CAR-T cells. In certain embodiments, complete response or better is determined less than 6 months after administration of CAR-T cells. In certain embodiments, complete response or better is determined less than 9 months after administration of CAR-T cells. In certain embodiments, complete response or better is determined less than 12 months after administration of CAR-T cells. In certain embodiments, complete response or better is determined less than 15 months after administration of CAR-T cells. In certain embodiments, complete response or better is determined more than 15 months after administration of CAR-T cells.

[0234] In certain embodiments, this treatment method is effective in achieving a best partial response or better in more than 80% of subjects. In certain embodiments, this treatment method is effective in achieving a best partial response or better in more than 85% of subjects. In certain embodiments, this treatment method is effective in achieving a best partial response or better in more than 86% of subjects. In certain embodiments, this treatment method is effective in achieving a best partial response or better in more than 87% of subjects. In certain embodiments, this treatment method is effective in achieving a best partial response or better in more than 88% of subjects. In certain embodiments, this treatment method is effective in achieving a best partial response or better in more than 89% of subjects. In certain embodiments, this treatment method is effective in achieving a best partial response or better in more than 90% of subjects. In certain embodiments, this treatment method is effective in achieving a best partial response or better in more than 91% of subjects. In certain embodiments, this treatment method is effective in achieving a best partial response or better in more than 92% of subjects. In certain embodiments, a best partial response or better is determined less than one month after administration of CAR-T cells. In certain embodiments, a best partial response or better is determined less than three months after administration of CAR-T cells. In certain embodiments, a best partial response or better is determined less than six months after administration of CAR-T cells. In certain embodiments, a best partial response or better is determined less than nine months after administration of CAR-T cells. In certain embodiments, a best partial response or better is determined less than twelve months after administration of CAR-T cells. In certain embodiments, a best partial response or better is determined less than fifteen months after administration of CAR-T cells. In certain embodiments, a best partial response or better is determined more than fifteen months after administration of CAR-T cells.

[0235] In certain embodiments, this treatment method is effective in achieving a median time to first response of less than 1.15 months. In certain embodiments, this treatment method is effective in achieving a median time to first response of less than 1.10 months. In certain embodiments, this treatment method is effective in achieving a median time to first response of less than 1.05 months. In certain embodiments, this treatment method is effective in achieving a median time to first response of less than 1.00 months. In certain embodiments, this treatment method is effective in achieving a median time to first response of less than 0.95 months.

[0236] In certain embodiments, this treatment method is effective in achieving a median time to best response of less than 2.96 months. In certain embodiments, this treatment method is effective in achieving a median time to best response of less than 2.86 months. In certain embodiments, this treatment method is effective in achieving a median time to best response of less than 2.76 months. In certain embodiments, this treatment method is effective in achieving a median time to best response of less than 2.66 months. In certain embodiments, this treatment method is effective in achieving a median time to best response of less than 2.56 months.

[0237] In certain embodiments, this method is effective in achieving an overall survival rate of over 80% nine months after CAR-T cell administration. In certain embodiments, this method is effective in achieving an overall survival rate of over 82% nine months after CAR-T cell administration. In certain embodiments, this method is effective in achieving an overall survival rate of over 85% nine months after CAR-T cell administration. In certain embodiments, this method is effective in achieving an overall survival rate of over 87% nine months after CAR-T cell administration. In certain embodiments, this method is effective in achieving an overall survival rate of over 90% nine months after CAR-T cell administration. In certain embodiments, this method is effective in achieving an overall survival rate of over 92% nine months after CAR-T cell administration. In certain embodiments, this method is effective in achieving an overall survival rate of over 95% nine months after CAR-T cell administration.

[0238] In certain embodiments, this method is effective in achieving an overall survival rate of over 80% 12 months after CAR-T cell administration. In certain embodiments, this method is effective in achieving an overall survival rate of over 83% 12 months after CAR-T cell administration. In certain embodiments, this method is effective in achieving an overall survival rate of over 86% 12 months after CAR-T cell administration. In certain embodiments, this method is effective in achieving an overall survival rate of over 89% 12 months after CAR-T cell administration. In certain embodiments, this method is effective in achieving an overall survival rate of over 92% 12 months after CAR-T cell administration. In certain embodiments, this method is effective in achieving an overall survival rate of over 93% 12 months after CAR-T cell administration.

[0239] In certain embodiments, this method is effective in achieving a progression-free survival rate of over 70% nine months after CAR-T cell administration. In certain embodiments, this method is effective in achieving a progression-free survival rate of over 72% nine months after CAR-T cell administration. In certain embodiments, this method is effective in achieving a progression-free survival rate of over 75% nine months after CAR-T cell administration. In certain embodiments, this method is effective in achieving a progression-free survival rate of over 77% nine months after CAR-T cell administration. In certain embodiments, this method is effective in achieving a progression-free survival rate of over 80% nine months after CAR-T cell administration. In certain embodiments, this method is effective in achieving a progression-free survival rate of over 82% nine months after CAR-T cell administration. In certain embodiments, this method is effective in achieving a progression-free survival rate of over 85% nine months after CAR-T cell administration. In certain embodiments, this method is effective in achieving a progression-free survival rate of 87% or higher 9 months after administration of CAR-T cells.

[0240] In certain embodiments, this method is effective in achieving a progression-free survival rate of over 66% 12 months after CAR-T cell administration. In certain embodiments, this method is effective in achieving a progression-free survival rate of over 69% 12 months after CAR-T cell administration. In certain embodiments, this method is effective in achieving a progression-free survival rate of over 72% 12 months after CAR-T cell administration. In certain embodiments, this method is effective in achieving a progression-free survival rate of over 76% 12 months after CAR-T cell administration. In certain embodiments, this method is effective in achieving a progression-free survival rate of over 80% 12 months after CAR-T cell administration. In certain embodiments, this method is effective in achieving a progression-free survival rate of over 84% 12 months after CAR-T cell administration.

[0241] In certain embodiments, this treatment method is effective in achieving recovery from cytokine release syndrome in more than 86% of subjects. In certain embodiments, this treatment method is effective in achieving recovery from cytokine release syndrome in more than 88% of subjects. In certain embodiments, this treatment method is effective in achieving recovery from cytokine release syndrome in more than 90% of subjects. In certain embodiments, this treatment method is effective in achieving recovery from cytokine release syndrome in more than 92% of subjects. In certain embodiments, this treatment method is effective in achieving recovery from cytokine release syndrome in more than 94% of subjects. In certain embodiments, this treatment method is effective in achieving recovery from cytokine release syndrome in more than 96% of subjects. In certain embodiments, this treatment method is effective in achieving recovery from cytokine release syndrome in more than 98% of subjects. In certain embodiments, this treatment method is effective in achieving recovery from cytokine release syndrome in more than 99% of subjects. In certain embodiments, this treatment method is effective in achieving recovery from cytokine release syndrome in 100% of subjects.

[0242] In certain embodiments, this treatment method is effective in achieving recovery in over 90% of subjects with immune effector cell-related neurotoxicity. In certain embodiments, this treatment method is effective in achieving recovery in over 92% of subjects with immune effector cell-related neurotoxicity. In certain embodiments, this treatment method is effective in achieving recovery in over 94% of subjects with immune effector cell-related neurotoxicity. In certain embodiments, this treatment method is effective in achieving recovery in over 96% of subjects with immune effector cell-related neurotoxicity. In certain embodiments, this treatment method is effective in achieving recovery in over 98% of subjects with immune effector cell-related neurotoxicity. In certain embodiments, this treatment method is effective in achieving recovery in 100% of subjects with immune effector cell-related neurotoxicity.

[0243] In some embodiments, the method further includes diagnosing the subject with respect to cytopenia. In some embodiments, cytopenia includes one or more, or all, of lymphopenia, neutropenia, and thrombocytopenia. Although not bound by theory, lymphopenia of grade 3 or grade 4 and not of grade 2 or lower is 0.5 × 10 per liter of the subject's blood sample. 9Neutropenia, characterized by a lymphocyte count of less than 1,000 cells, is characterized by a neutrophil count of less than 1,000 cells per microliter of blood sample, and thrombocytopenia, characterized by a platelet count of less than 50,000 cells per microliter of blood sample. In some embodiments, more than 75% of subjects with grade 3 or grade 4 lymphopenia after CAR-T cell administration recover to grade 2 or less lymphopenia 60 days after CAR-T cell administration. In some embodiments, more than 80% of subjects with grade 3 or grade 4 lymphopenia after CAR-T cell administration recover to grade 2 or less lymphopenia 60 days after CAR-T cell administration. In some embodiments, more than 85% of subjects with grade 3 or grade 4 lymphopenia after CAR-T cell administration recover to grade 2 or less lymphopenia 60 days after CAR-T cell administration. In some embodiments, more than 90% of subjects with grade 3 or grade 4 lymphopenia after CAR-T cell administration recover to grade 2 or lower lymphopenia 60 days after CAR-T cell administration. In some embodiments, more than 70% of subjects with grade 3 or grade 4 neutropenia after CAR-T cell administration recover to grade 2 or lower neutropenia 60 days after CAR-T cell administration. In some embodiments, more than 75% of subjects with grade 3 or grade 4 neutropenia after CAR-T cell administration recover to grade 2 or lower neutropenia 60 days after CAR-T cell administration. In some embodiments, more than 80% of subjects with grade 3 or grade 4 neutropenia after CAR-T cell administration recover to grade 2 or lower neutropenia 60 days after CAR-T cell administration. In some embodiments, more than 85% of subjects with grade 3 or grade 4 neutropenia after CAR-T cell administration recover to grade 2 or lower neutropenia 60 days after CAR-T cell administration.In some embodiments, more than 30% of subjects with grade 3 or grade 4 thrombocytopenia after CAR-T cell administration recover to grade 2 or lower thrombocytopenia 60 days after CAR-T cell administration. In some embodiments, more than 34% of subjects with grade 3 or grade 4 thrombocytopenia after CAR-T cell administration recover to grade 2 or lower thrombocytopenia 60 days after CAR-T cell administration. In some embodiments, more than 38% of subjects with grade 3 or grade 4 thrombocytopenia after CAR-T cell administration recover to grade 2 or lower thrombocytopenia 60 days after CAR-T cell administration. In some embodiments, more than 42% of subjects with grade 3 or grade 4 thrombocytopenia after CAR-T cell administration recover to grade 2 or lower thrombocytopenia 60 days after CAR-T cell administration.

[0244] In certain embodiments, the subject is re-treated by a second intravenous infusion of a second dose of CAR-T cells. In certain embodiments, the re-treatment dose is 1.0 × 10⁶ per kilogram of the subject's body weight. 5 ~5.0×10 6 Contains 10 CAR-T cells. In a particular embodiment, the retreatment dose is approximately 0.75 × 10¹⁶ per kilogram of the subject's body weight. 5 Contains 1 CAR-T cell. In certain embodiments, the subject is retreated when disease progression occurs after the best response, above minimum response, is achieved following the initial infusion of CAR-T cells. In certain embodiments, the time between the initial infusion of CAR-T cells and the detection of disease progression includes at least 6 months.

[0245] Manufacturing kits and articles Any of the compositions described herein may be included in the kit. In some embodiments, the kit provides modified immortalized CAR-T cells, which may also include reagents suitable for cell expansion, such as a culture medium.

[0246] In non-limiting examples, a chimeric receptor expression construct, one or more reagents for constructing a chimeric receptor expression construct, cells for transfection of the expression construct, and / or one or more instruments for obtaining immortalized T cells for transfection of the expression construct (such instruments may be syringes, pipettes, forceps, and / or any such medically approved instruments).

[0247] In some embodiments, the kit includes reagents or equipment for electroporation of cells.

[0248] In some embodiments, the kit includes artificial antigen-presenting cells.

[0249] A kit may comprise one or more suitably divided compositions of the Disclosure or reagents for preparing compositions of the Disclosure. The components of the kit may be packaged in either aqueous medium or lyophilized form. The container means of the kit may comprise at least one vial, test tube, flask, bottle, syringe, or other container means in which the components may be placed, preferably suitably divided. If there are two or more components in the kit, the kit will also generally comprise a second, third, or other additional container in which additional components may be placed separately. However, various combinations of components may be contained in vials. The kits of the Disclosure will also typically comprise means for tightly sealing and containing the chimeric receptor construct and any other reagent containers for commercial sale. Such containers may comprise, for example, injection-molded or blow-molded plastic containers in which the desired vials are held.

[0250] The following exemplary embodiments are intended merely to illustrate the Disclosure and should not be construed as limiting the Disclosure in any way. Exemplary Embodiments 1. A method of treating the subject, (a) Extracellular antigen-binding domain having specific binding ability to B cell maturation antigen (BCMA) epitopes, (b) Transmembrane domain, and (c) Intracellular signaling domain A method comprising administering a certain dose of chimeric antigen receptor (CAR)-containing T cells, The study involves patients with multiple myeloma who have received 1 to 3 prior treatment lines, including immunomodulatory drug (IMiD) therapy, and who are resistant to IMiD. 2. The method according to Embodiment 1, wherein the subject has high-risk features, and, at random selection, the high-risk features are cytogenetic abnormalities, International Staging Classification (ISS) stage III, and / or soft tissue plasmacytoma. 3. A method of selectively treating the target, (1) Determining whether the subject has high-risk features, wherein the high-risk features are cytogenetic abnormalities, International Staging Classification (ISS) stage III, and / or soft tissue plasmacytoma; and (2) For subjects determined to have high-risk characteristics in step (1), (a) Extracellular antigen-binding domain having specific binding ability to BCMA epitopes, (b) Transmembrane domain, and (c) Intracellular signaling domain Administering a certain dose of chimeric antigen receptor (CAR)-containing T cells, including In a method including, The method involves electively selecting subjects who have multiple myeloma, have received 1 to 3 prior treatment lines, including IMiD therapy, and are resistant to IMiD. 4. A method for selectively treating subjects, wherein subjects determined to have high-risk characteristics, (a) Extracellular antigen-binding domain having specific binding ability to BCMA epitopes, (b) Transmembrane domain, and (c) Intracellular signaling domain A method comprising administering a certain dose of chimeric antigen receptor (CAR)-containing T cells, High-risk features include cytogenetic abnormalities, International Staging System (ISS) stage III, and / or soft tissue plasmacytoma. The method involves electively selecting subjects who have multiple myeloma, have received 1 to 3 prior treatment lines, including IMiD therapy, and are resistant to IMiD. 5. The method according to any one of Embodiments 1 to 4, wherein IMiD is lenalidomide. 6. The method according to any one of Embodiments 2 to 5, wherein the high-risk feature is a cytogenetic abnormality. 7. The method according to Embodiment 6, wherein the cytogenetic abnormality is a high-risk cytogenetic abnormality. 8. The method according to Embodiment 7, wherein the subject has one or more high-risk cytogenetic abnormalities selected from the group including Gain / amp(1q), del(17p), t(4;14), t(14;16), or any combination thereof. 9. The method according to Embodiment 8, wherein the cytogenetic abnormality includes Gain / amp(1q). 10. The method according to Embodiment 8, wherein the cytogenetic abnormality includes del(17p). 11. The method according to Embodiment 8, wherein the cytogenetic abnormality includes t(4;14). 12. The method according to Embodiment 8, wherein the cytogenetic abnormality includes t(14;16). 13. The method according to any one of Embodiments 6 to 12, wherein the subject has at least two cytogenetic abnormalities, and optionally the subject has two, three, four, five, or more cytogenetic abnormalities. 14. The method according to Embodiment 6, wherein the cytogenetic abnormality is a standard-risk cytogenetic abnormality. 15. The method according to any one of Embodiments 2 to 5, wherein the high-risk feature is International Staging Classification (ISS) Stage III. 16. The method according to any one of Embodiments 2 to 5, wherein the high-risk feature is soft tissue plasmacytoma. 17. The method according to any one of Embodiments 1 to 16, wherein the subject has received one prior treatment line. 18. The method according to any one of Embodiments 1 to 16, wherein the subject has received two prior treatment lines. 19. The method according to any one of Embodiments 1 to 16, wherein the subject has received three prior treatment lines. 20.1, The method according to any one of Embodiments 1 to 19, wherein two or three prior treatment lines include treatment with pomalidomide. 21.1, The method according to any one of Embodiments 1 to 20, wherein two or three prior treatment lines further comprise treatment with an anti-CD38 antibody, and optionally the anti-CD38 antibody is daratumumab and / or isatuximab. 22.1, The method according to any one of Embodiments 1 to 21, wherein two or three prior treatment lines further comprise treatment with a proteasome inhibitor, and optionally the proteasome inhibitor is bortezomib, carfilzomib, ixazomib, or any combination thereof. 23. The method according to any one of Embodiments 1 to 22, wherein the subject has previously received bridging therapy, and voluntarily, the bridging therapy is at the physician's discretion, and voluntarily, the bridging therapy comprises pomalidomide, bortezomib, dexamethasone, daratumumab, or any combination thereof, and further voluntarily, the bridging therapy comprises pomalidomide, bortezomib, and dexamethasone, and further voluntarily, the bridging therapy comprises daratumumab, pomalidomide, and dexamethasone. 24. The method according to Embodiment 23, wherein the subject has received bridging therapy approximately every 20 to 30 days, optionally, has received bridging therapy approximately every 21 days, optionally, has received bridging therapy approximately every 28 days, and optionally, has received at least one, two, three, four, or more bridging therapies. 25. The subjects had previously received lymphocyte depletion therapy, and optionally, the lymphocyte depletion therapy included daily cyclophosphamide and / or fludarabine, and optionally, the lymphocyte depletion therapy included daily cyclophosphamide and fludarabine, and optionally, the lymphocyte depletion therapy included approximately 300 mg / m² daily for 3 days.2 The concentration of cyclophosphamide is approximately 30 mg / m². 2 The method according to any one of Embodiments 1 to 24, comprising fludarabine at the specified concentration. 26. The dose of T cells is 0.5-1.0 × 10⁻⁶ 6 The cell-to-subject ratio is 1 kg, and selection is random, with a T cell dose of approximately 0.75 × 10⁶ 6 The method according to any one of Embodiments 1 to 25, wherein the cell / subject body weight is 1 kg, and optionally, the method includes administering a dose of T cells approximately 5 to 7 days after the start of lymphocyte depletion therapy, and optionally, the dose is administered as a single infusion. 27. The method according to any one of Embodiments 1 to 26, which is effective in achieving a total response in a subject after administering a dose of T cells to the subject, is optionally effective in achieving a total response rate of approximately 75% to approximately 100%, is optionally effective in achieving a total response rate of approximately 84.6%, and is optionally effective in achieving a total response rate of approximately 99.4%. 28. The overall response rate is ranked from best to worst. (1) Strict complete response; (2) Complete response; (3) Best partial response; (4) Partial response; or (5) Minimum response The method according to Embodiment 27, including the method described in Embodiment 27. 29. The method according to Embodiment 28, wherein the overall response is defined as a strict complete response, and is effective in achieving strict complete responses at a rate of approximately 40% to approximately 90%, approximately 50% to approximately 80%, approximately 58.2%, or approximately 68.8%, at any discretion. 30. The method according to Embodiment 28, wherein all responses are complete responses, and the method is optionally effective in achieving complete responses in a ratio of approximately 10% to approximately 20%, and optionally effective in achieving complete responses in a ratio of approximately 14.9% or approximately 17.6%. 31. The method according to Embodiment 28, wherein the total response is the best partial response or a partial response. 32. (1) Effective in achieving strict complete response or complete response in approximately 70% to 90% of cases, and, optionally, effective in achieving strict complete response or complete response in approximately 73.1% or 86.4% of cases; (2) Effective in achieving strict complete response, complete response, or best partial response in approximately 80% to 100% of cases, and optionally effective in achieving strict complete response, complete response, or best partial response in approximately 81.3% or 96.0% of cases; (3) Effective in achieving minimal response; (4) Effective in further achieving minimal residual disease negativity, and optionally effective in achieving minimal residual disease negativity in approximately 50% to 80% of cases, and optionally effective in achieving minimal residual disease negativity in approximately 60.6% or 71.6% of cases; or (5) At least approximately 60-100% of subjects, at least approximately 69.4-81.1% of subjects, or at least approximately 84.1-93.4% of subjects are able to achieve further 12-month progression-free survival, and on a voluntary basis, at least approximately 75.9% of subjects or approximately 89.7% of subjects are able to achieve 12-month progression-free survival. The method described in Embodiment 28. 33. (1) The time to complete response or minimum response on the first attempt is in the range of approximately 0.9 to approximately 11.1 months, and optionally, the time to complete response or minimum response on the first attempt is approximately 2.1 months on a median basis; or (2) The time to best overall response or best minimum response is approximately 1.1 to 18.6 months, and for optional selection, the median time to best overall response or best minimum response is approximately 6.4 or 6.5 months. The method according to any one of embodiments 27 to 32. 34. The method further comprises treating the subject for adverse events after administering a dose of T cells, optionally comprising administering a treatment to the subject to mitigate the adverse events, optionally comprising the adverse events being hematological adverse events, non-hematological adverse events, adverse events occurring under study treatment, or any combination thereof, optionally comprising the non-hematological adverse events being infections and / or non-hematological adverse events other than infections, and optionally comprising the adverse events being neutropenia, thrombocytopenia, anemia, lymphopenia, upper respiratory tract infection, nasopharyngitis, sinusitis, rhinitis, tonsillitis, pharyngitis, laryngitis, pharyngotonsillitis, COVID-19, COVID-19 pneumonia, asymptomatic COVID-19, neutropenic sepsis, progressive multifocal leukoencephalopathy The method according to any one of Embodiments 1 to 33, comprising any adverse event including leukoencephalpathy, septic shock, respiratory failure, pulmonary embolism, lower respiratory tract / lung infection, pneumonia, bronchitis, nausea, hypogammaglobulinemia, diarrhea, fatigue, headache, constipation, hypokalemia, asthenia, peripheral edema, decreased appetite, peripheral sensory neuropathy, back pain, arthralgia, fever, dyspnea, insomnia, or any combination thereof, further optionally, the adverse event is a Grade 3 / 4 adverse event, and further optionally, the adverse event persists for more than about 30 days or about 60 days. 35. The method according to any one of Embodiments 1 to 34, wherein the method further comprises treating a subject with respect to a secondary primary malignancy after administering a dose of T cells, optionally comprising administering a treatment to the subject to alleviate the secondary primary malignancy, optionally comprising a cutaneous / non-invasive malignancy, a hematological malignancy, a non-cutaneous / invasive malignancy, or any combination thereof, and optionally comprising a basal cell carcinoma, Bowen's disease, squamous cell carcinoma of the lip, malignant melanoma, malignant melanoma in situ, squamous cell carcinoma of the skin, acute myeloid leukemia, myelodysplastic syndrome, peripheral T-cell lymphoma, angiosarcoma, invasive lobular breast carcinoma, pleomorphic malignant fibrous histiocytoma, renal cell carcinoma, tonsil carcinoma, or any combination thereof. 36. The method according to Embodiment 34 or 35, wherein adverse events or secondary primary malignancies occur in the subject at a rate equivalent to that of subjects receiving standard treatment. 37. The method further comprises treating the subject for CAR-T related adverse events after administering a dose of T cells, optionally comprising administering treatment to the subject to mitigate CAR-T related adverse events, optionally comprising the CAR-T related adverse events including cytokine release syndrome (CRS) and / or neurotoxicity, optionally (a) The CAR-T related adverse event is CRS, and optionally, CRS occurs in approximately 60% to 90% or 76.1% of the subjects, and optionally, the maximum toxicity grade of CRS is Grade 1, Grade 2 or Grade 3, and optionally, (1) The maximum toxicity grade of CRS is Grade 1, and, in optional selection, the maximum toxicity grade of Grade 1 occurs in approximately 52.8% of the subjects; (2) The maximum toxicity grade of CRS is Grade 2, and, on an optional basis, the maximum toxicity grade of Grade 2 occurs in approximately 22.2% of the subjects; (3) The maximum toxicity grade of CRS is Grade 3, and, on an optional basis, the maximum toxicity grade of Grade 3 occurs in approximately 1.1% of the subjects; (4) The time to the first onset of CRS is in the range of approximately 1 to 23 days, and, on a random selection basis, the median time to the first onset of CRS is approximately 8 days; (5) The duration of CRS is in the range of approximately 1 to approximately 17 days, and optionally, the median duration of CRS is approximately 3 days; or (6) Treatment includes tocilizumab, oxygen, corticosteroids, vasopressors, or any combination thereof; or (b) The CAR-T related adverse event is neurotoxic, and optionally the neurotoxicity includes immunoeffector cell-associated neurotoxic syndrome or associated symptoms, motor / neurocognitive neurotoxicity, neurotoxic adverse events occurring under study treatment, non-immune effector cell-associated neurotoxic syndrome or associated symptoms, or any combination thereof, and optionally the neurotoxicity is immunoeffector cell-associated neurotoxic syndrome or associated symptoms, and optionally the (1) Immunoeffector cell-associated neurotoxicity syndrome or associated symptoms occur in approximately 4.5% of the subjects; (2) The maximum toxicity grade of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is Grade 1 or Grade 2, and optionally, the maximum toxicity grade of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is Grade 1, and optionally, Grade 1 maximum toxicity occurs in approximately 3.4% of the subjects, or optionally, the maximum toxicity grade of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is Grade 2, and optionally, Grade 2 maximum toxicity occurs in approximately 1.1% of the subjects; (3) The time to onset of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is in the range of approximately 6 to 15 days, and, on an optional basis, the median time to onset of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is approximately 9.5 days; (4) The duration of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is in the range of approximately 1 to approximately 6 days, and optionally, the median duration of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is approximately 2 days; or (5) Treatment including corticosteroids and / or tocilizumab The method according to any one of Embodiments 1 to 36. 38. Neurotoxicity is CAR-T cell neurotoxicity, and, at random selection, CAR-T cell neurotoxicity occurs in approximately 17.0% of subjects, and, at random selection, CAR-T cell neurotoxicity includes grade 3 / 4 neurotoxicity, grade 5 neurotoxicity, cranial nerve palsy, peripheral neuropathy, adverse events related to motor / neurocognitive function that occurred during the study treatment, or any combination thereof, and further at random selection, (a) CAR-T cell neurotoxicity is a grade 3 / 4 neurotoxicity occurring in approximately 2.3% of the subjects; (b) CAR-T cell neurotoxicity is grade 5 neurotoxicity; (c) CAR-T cell neurotoxicity is cranial nerve palsy, and in random selection, cranial nerve palsy occurs in approximately 9.1% of the subjects, and in random selection, (1) The cranial nerve palsy is of grade 2 or grade 3, and further, on an optional basis, the cranial nerve palsy is of grade 2, occurring in approximately 8.0% of the subjects, and further, on an optional basis, the cranial nerve palsy is of grade 3, occurring in approximately 1.1% of the subjects; (2) The time from administration of T-cells to the onset of cranial nerve palsy in subjects ranges from approximately 17 to 60 days, and in a randomly selected group, the median time from administration of T-cells to the onset of cranial nerve palsy in subjects is approximately 21 days; (3) Cranial nerve palsy occurs in the third, fifth, or seventh cranial nerve; (4) The duration of cranial nerve palsy is in the range of approximately 15 to 262 days, and optionally, the median duration of cranial nerve palsy is approximately 77 days; or (5) Treatment may include corticosteroids; (d) CAR-T cell neurotoxicity is peripheral neuropathy, and peripheral neuropathy occurs in approximately 2.8% of subjects on an optional basis; or (e) CAR-T cell neurotoxicity is an adverse event that occurs during the study treatment for motor and neurocognitive function, and the adverse event that occurs during the study treatment for motor and neurocognitive function is of grade 1, and further randomly selected, the proportion of subjects experiencing a grade 1 adverse event during the study treatment for motor and neurocognitive function is approximately 0.6%. The method according to Embodiment 37. 39.(a) CD3+ cells containing CAR in the subject's blood reach a peak approximately 13 days after T cell administration to the subject, and, in optional selection, CD3+ cells containing CAR in the subject's blood reach a peak at an average concentration of approximately 1523 cells / μL; (b) CD3+ cells containing CAR in the subject's blood remain detectable approximately 13 to 631 days after T cell administration to the subject, and optionally, CD3+ cells containing CAR in the subject's blood remain detectable approximately 57 days after T cell administration to the subject; or (c) AUC of CD3+ cells containing CAR in the blood of the subject 0-28 However, the average value is approximately 12,504 cells / μL. The method according to any one of Embodiments 1 to 38. 40. A first VHH domain containing CDR1, CDR2, and CDR3 as shown in the VHH domain containing the amino acid sequence of SEQ ID NO: 2, a second VHH domain containing CDR1, CDR2, and CDR3 as shown in the VHH domain containing the amino acid sequence of SEQ ID NO: 4, optionally, the first VHH domain containing CDR1 containing the amino acid sequence of SEQ ID NO: 18, CDR2 containing the amino acid sequence of SEQ ID NO: 19, and CDR3 containing the amino acid sequence of SEQ ID NO: 20, and the second The VHH domain includes CDR1 containing the amino acid sequence of SEQ ID NO: 21, CDR2 containing the amino acid sequence of SEQ ID NO: 22, and CDR3 containing the amino acid sequence of SEQ ID NO: 23, and optionally, the first VHH domain contains the amino acid sequence of SEQ ID NO: 2, and the second VHH domain contains the amino acid sequence of SEQ ID NO: 4, and optionally, the first VHH domain is at the N-terminus of the second VHH domain, or the first VHH domain is at the C-terminus of the second VHH domain, and optionally, (a) The first VHH domain is linked to the second VHH domain by a linker containing the amino acid sequence of SEQ ID NO: 3; (b) The transmembrane domain is derived from a molecule selected from the group consisting of CD8α, CD4, CD28, CD137, CD80, CD86, CD152, and PD1, and optionally the transmembrane domain is derived from CD8α and contains the amino acid sequence of SEQ ID NO: 6; (c) The intracellular signaling domain includes the primary intracellular signaling domain of an immune effector cell, and optionally, the primary intracellular signaling domain is derived from CD3ζ containing the amino acid sequence of SEQ ID NO: 8; (d) The intracellular signaling domain includes a co-stimulatory signaling domain, which optionally derives from a co-stimulatory molecule selected from the group consisting of ligands for CD27, CD28, CD137, OX40, CD30, CD40, CD3, LFA-1, ICOS, CD2, CD7, LIGHT, NKG2C, B7-H3, CD83 and any combination thereof, and optionally derives from a co-stimulatory signaling domain including the cytoplasmic domain of CD137 containing the amino acid sequence of SEQ ID NO: 7; (e) The CAR further comprises a hinge domain located between the C-terminus of the extracellular antigen-binding domain and the N-terminus of the transmembrane domain, and optionally, the hinge domain is derived from CD8α containing the amino acid sequence of SEQ ID NO: 5; (f) The CAR further comprises a signal peptide located at the N-terminus of the polypeptide, and optionally the signal peptide is derived from CD8α having the amino acid sequence of SEQ ID NO: 1; or (g)CAR contains the amino acid sequence of SEQ ID NO: 17, The method according to any one of Embodiments 1 to 39. 41. The method according to any one of Embodiments 1 to 40, wherein the dose of T cells is formulated in a composition containing 5% dimethyl sulfoxide (DMSO). 42. The method according to any one of Embodiments 1 to 41, wherein administering a dose of T cells reduces the risk of disease progression or death in the subject. 43. The method according to Embodiment 42, wherein the risk of disease progression or death is reduced compared to the administration of daratumumab-pomalidomide-dexamethasone (DPd) or pomalidomide-bortezomib-dexamethasone (PVd) therapy. 44. The method according to Embodiment 42, wherein the risk of disease progression or death is reduced compared to the administration of ide-cel therapy. 45. The method according to any one of Embodiments 42 to 44, wherein the subject exhibits a disease progression or mortality risk reduced by approximately 60% to approximately 75%. 46. ​​The method according to Embodiment 45, wherein the subject exhibits a disease progression or mortality risk reduced by approximately 74%. 47. The method according to any one of embodiments 42 to 46, wherein IMiD is lenalidomide. 48. The method according to any one of embodiments 42 to 47, wherein the subject has received three or fewer prior treatment lines. 49. The method according to any one of embodiments 42 to 47, wherein the subject has received two or fewer prior treatment lines. 50. The method according to any one of embodiments 42 to 47, wherein the subject has received only one prior treatment line. 51. The method according to any one of embodiments 42 to 50, which is effective in achieving an overall response rate (ORR) of approximately 75% to approximately 100%. 52. The method according to Embodiment 51, wherein the ORR is approximately 84.6%. 53. The method according to any one of Embodiments 42 to 52, wherein the treatment is effective in extending the median progression-free survival (PFS) of the subject compared to administration of DPd or PVd treatment. 54. The method according to Embodiment 53, wherein the PFS at 12 months after administration of the treatment is approximately 75.9%. 55. The method according to Embodiment 54, wherein the PFS at 12 months after administration of DPd or PVd is approximately 48.6%. 56. The method according to any one of Embodiments 1 to 23 and 42 to 55, wherein the treatment is more effective in achieving a strict complete response (sCR) in the subject compared to the administration of DPd or PVd treatment. 57. The method according to embodiment 56, wherein the sCR after administration of the treatment is approximately 58.2%. 58. The method according to Embodiment 57, wherein the sCR after administration of DPd or PVd is approximately 15.2%. 59. The method according to any one of Embodiments 1 to 23 and 42 to 58, wherein the treatment is more effective in achieving best partial response (VGPR) or better in the subject compared to administration of DPd or PVd treatment. 60. The method according to Embodiment 59, wherein approximately 81.3% of patients achieve a response of VGPR or better after administration of the treatment. 61. The method according to Embodiment 60, wherein approximately 45.5% of patients achieve a response of VGPR or better after administration of DPd or PVd. 62. The method further comprises treating the subject for CAR-T related adverse events after administering a dose of T cells, optionally comprising administering treatment to the subject to mitigate CAR-T related adverse events, optionally comprising the CAR-T related adverse events including cytokine release syndrome (CRS) and / or neurotoxicity, optionally (a) The CAR-T related adverse event is CRS, and optionally, CRS occurs in approximately 60% to 90% or 76.1% of the subjects, and optionally, the maximum toxicity grade of CRS is Grade 1, Grade 2 or Grade 3, and optionally, (1) The maximum toxicity grade of CRS is Grade 1, and, in optional selection, the maximum toxicity grade of Grade 1 occurs in approximately 52.8% of the subjects; (2) The maximum toxicity grade of CRS is Grade 2, and, on an optional basis, the maximum toxicity grade of Grade 2 occurs in approximately 22.2% of the subjects; (3) The maximum toxicity grade of CRS is Grade 3, and, on an optional basis, the maximum toxicity grade of Grade 3 occurs in approximately 1.1% of the subjects; (4) The time to the first onset of CRS is in the range of approximately 1 to 23 days, and, on a random selection basis, the median time to the first onset of CRS is approximately 8 days; (5) The duration of CRS is in the range of approximately 1 to approximately 17 days, and optionally, the median duration of CRS is approximately 3 days; or (6) Treatment includes tocilizumab, oxygen, corticosteroids, vasopressors, or any combination thereof; or (b) The CAR-T related adverse event is neurotoxic, and optionally the neurotoxicity includes immunoeffector cell-associated neurotoxic syndrome or associated symptoms, motor / neurocognitive neurotoxicity, neurotoxic adverse events occurring under study treatment, non-immune effector cell-associated neurotoxic syndrome or associated symptoms, or any combination thereof, and optionally the neurotoxicity is immunoeffector cell-associated neurotoxic syndrome or associated symptoms, and optionally the (1) Immunoeffector cell-associated neurotoxicity syndrome or associated symptoms occur in approximately 4.5% of the subjects; (2) The maximum toxicity grade of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is Grade 1 or Grade 2, and optionally, the maximum toxicity grade of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is Grade 1, and optionally, Grade 1 maximum toxicity occurs in approximately 3.4% of the subjects, or optionally, the maximum toxicity grade of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is Grade 2, and optionally, Grade 2 maximum toxicity occurs in approximately 1.1% of the subjects; (3) The time to onset of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is in the range of approximately 6 to 15 days, and, on an optional basis, the median time to onset of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is approximately 9.5 days; (4) The duration of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is in the range of approximately 1 to approximately 6 days, and optionally, the median duration of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is approximately 2 days; or (5) Treatment including corticosteroids and / or tocilizumab The method according to any one of embodiments 42 to 61. 63. Neurotoxicity is CAR-T cell neurotoxicity, and in random selection, CAR-T cell neurotoxicity occurs in approximately 17.0% of subjects, and in random selection, CAR-T cell neurotoxicity includes grade 3 / 4 neurotoxicity, grade 5 neurotoxicity, cranial nerve palsy, peripheral neuropathy, adverse events related to motor / neurocognitive function that occurred during the study treatment, or any combination thereof, and further random selection, (a) CAR-T cell neurotoxicity is a grade 3 / 4 neurotoxicity occurring in approximately 2.3% of the subjects; (b) CAR-T cell neurotoxicity is grade 5 neurotoxicity; (c) CAR-T cell neurotoxicity is cranial nerve palsy, and in random selection, cranial nerve palsy occurs in approximately 9.1% of the subjects, and in random selection, (1) The cranial nerve palsy is of grade 2 or grade 3, and further, on an optional basis, the cranial nerve palsy is of grade 2, occurring in approximately 8.0% of the subjects, and further, on an optional basis, the cranial nerve palsy is of grade 3, occurring in approximately 1.1% of the subjects; (2) The time from administration of T-cells to the onset of cranial nerve palsy in subjects ranges from approximately 17 to 60 days, and in a randomly selected group, the median time from administration of T-cells to the onset of cranial nerve palsy in subjects is approximately 21 days; (3) Cranial nerve palsy occurs in the third, fifth, or seventh cranial nerve; (4) The duration of cranial nerve palsy is in the range of approximately 15 to 262 days, and optionally, the median duration of cranial nerve palsy is approximately 77 days; or (5) Treatment may include corticosteroids; (d) CAR-T cell neurotoxicity is peripheral neuropathy, and peripheral neuropathy occurs in approximately 2.8% of subjects on an optional basis; or (e) CAR-T cell neurotoxicity is an adverse event that occurs during the study treatment for motor and neurocognitive function, and the adverse event that occurs during the study treatment for motor and neurocognitive function is of grade 1, and further randomly selected, the proportion of subjects experiencing a grade 1 adverse event during the study treatment for motor and neurocognitive function is approximately 0.6%. The method according to Embodiment 62. 64. The method according to any one of embodiments 42 to 63, wherein the subject has one or more high-risk cytogenetic abnormalities selected from the group including Gain / amp(1q), del(17p), t(4;14), t(14;16), or any combination thereof. 65. The method according to Embodiment 64, wherein the subject has at least two cytogenetic abnormalities, and optionally the subject has two, three, four, five, or more cytogenetic abnormalities. 66. The method according to Embodiment 64 or 65, wherein the cytogenetic abnormality is a standard-risk cytogenetic abnormality. 67. The method according to any one of Embodiments 1 to 66, wherein the administration of a dose of T cells is more effective in achieving a better-than-best partial response (VGPR) or better in a subject compared to the administration of DPd or PVd therapy. 68. The method according to Embodiment 67, wherein approximately 81.3% of patients achieve a response of VGPR or better after administration of the treatment. 69. The method according to Embodiment 68, wherein approximately 45.5% of patients achieve a response of VGPR or better after administration of DPd or PVd. 70. The method according to any one of embodiments 67 to 69, wherein the treatment is more effective in achieving a strict complete response (sCR) in the subject compared to the administration of DPd or PVd treatment. 71. The method according to Embodiment 70, wherein the sCR after administration of the treatment is approximately 58.2%. 72. The method according to Embodiment 71, wherein the sCR after administration of DPd or PVd is approximately 15.2%. 73. The method according to any one of embodiments 67 to 72, wherein administering a dose of T cells reduces the risk of disease progression or death in the subject. 74. The method according to Embodiment 73, wherein the risk of disease progression or death is reduced compared to the administration of daratumumab-pomalidomide-dexamethasone (DPd) or pomalidomide-bortezomib-dexamethasone (PVd) therapy. 75. The method according to Embodiment 73, wherein the risk of disease progression or death is reduced compared to the administration of ide-cel therapy. 76. The method according to any one of embodiments 67 to 75, wherein the subject exhibits a disease progression or mortality risk reduced by approximately 60% to approximately 75%. 77. The method according to Embodiment 76, wherein the subject exhibits a disease progression or mortality risk reduced by approximately 74%. 78. The method according to any one of embodiments 67 to 77, wherein IMiD is lenalidomide. 79. The method according to any one of embodiments 67 to 78, wherein the subject has received three or fewer prior treatment lines. 80. The method according to any one of embodiments 67 to 79, wherein the subject has received two or fewer prior treatment lines. 81. The method according to any one of embodiments 67 to 80, wherein the subject has received only one prior treatment line. 82. The method according to any one of embodiments 67 to 81, which is effective in achieving an overall response rate (ORR) of approximately 75% to approximately 100%. 83. The method according to Embodiment 82, wherein the ORR is approximately 84.6%. 84. The method according to any one of Embodiments 67 to 83, wherein the treatment is effective in extending the median progression-free survival (PFS) of the subject compared to administration of DPd or PVd treatment. 85. The method according to Embodiment 84, wherein the PFS at 12 months after administration of treatment is approximately 75.9%. 86. The method according to Embodiment 85, wherein the PFS at 12 months after administration of DPd or PVd is approximately 48.6%. 87. The method further comprises treating the subject for CAR-T related adverse events after administering a dose of T cells, optionally comprising administering treatment to the subject to mitigate CAR-T related adverse events, optionally comprising the CAR-T related adverse events including cytokine release syndrome (CRS) and / or neurotoxicity, optionally (a) The CAR-T related adverse event is CRS, and optionally, CRS occurs in approximately 60% to 90% or 76.1% of the subjects, and optionally, the maximum toxicity grade of CRS is Grade 1, Grade 2 or Grade 3, and optionally, (1) The maximum toxicity grade of CRS is Grade 1, and, in optional selection, the maximum toxicity grade of Grade 1 occurs in approximately 52.8% of the subjects; (2) The maximum toxicity grade of CRS is Grade 2, and, on an optional basis, the maximum toxicity grade of Grade 2 occurs in approximately 22.2% of the subjects; (3) The maximum toxicity grade of CRS is Grade 3, and, on an optional basis, the maximum toxicity grade of Grade 3 occurs in approximately 1.1% of the subjects; (4) The time to the first onset of CRS is in the range of approximately 1 to 23 days, and, on a random selection basis, the median time to the first onset of CRS is approximately 8 days; (5) The duration of CRS is in the range of approximately 1 to approximately 17 days, and optionally, the median duration of CRS is approximately 3 days; or (6) Treatment includes tocilizumab, oxygen, corticosteroids, vasopressors, or any combination thereof; or (b) The CAR-T related adverse event is neurotoxic, and optionally the neurotoxicity includes immunoeffector cell-associated neurotoxic syndrome or associated symptoms, motor / neurocognitive neurotoxicity, neurotoxic adverse events occurring under study treatment, non-immune effector cell-associated neurotoxic syndrome or associated symptoms, or any combination thereof, and optionally the neurotoxicity is immunoeffector cell-associated neurotoxic syndrome or associated symptoms, and optionally the (1) Immunoeffector cell-associated neurotoxicity syndrome or associated symptoms occur in approximately 4.5% of the subjects; (2) The maximum toxicity grade of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is Grade 1 or Grade 2, and optionally, the maximum toxicity grade of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is Grade 1, and optionally, Grade 1 maximum toxicity occurs in approximately 3.4% of the subjects, or optionally, the maximum toxicity grade of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is Grade 2, and optionally, Grade 2 maximum toxicity occurs in approximately 1.1% of the subjects; (3) The time to onset of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is in the range of approximately 6 to 15 days, and, on an optional basis, the median time to onset of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is approximately 9.5 days; (4) The duration of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is in the range of approximately 1 to approximately 6 days, and optionally, the median duration of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is approximately 2 days; or (5) Treatment including corticosteroids and / or tocilizumab The method according to any one of embodiments 67 to 86. 88. Neurotoxicity is CAR-T cell neurotoxicity, and, at random selection, CAR-T cell neurotoxicity occurs in approximately 17.0% of subjects, and, at random selection, CAR-T cell neurotoxicity includes grade 3 / 4 neurotoxicity, grade 5 neurotoxicity, cranial nerve palsy, peripheral neuropathy, adverse events related to motor / neurocognitive function that occurred during the study treatment, or any combination thereof, and further at random selection, (a) CAR-T cell neurotoxicity is a grade 3 / 4 neurotoxicity occurring in approximately 2.3% of the subjects; (b) CAR-T cell neurotoxicity is grade 5 neurotoxicity; (c) CAR-T cell neurotoxicity is cranial nerve palsy, and in random selection, cranial nerve palsy occurs in approximately 9.1% of the subjects, and in random selection, (1) The cranial nerve palsy is of grade 2 or grade 3, and further, on an optional basis, the cranial nerve palsy is of grade 2, occurring in approximately 8.0% of the subjects, and further, on an optional basis, the cranial nerve palsy is of grade 3, occurring in approximately 1.1% of the subjects; (2) The time from administration of T-cells to the onset of cranial nerve palsy in subjects ranges from approximately 17 to 60 days, and in a randomly selected group, the median time from administration of T-cells to the onset of cranial nerve palsy in subjects is approximately 21 days; (3) Cranial nerve palsy occurs in the third, fifth, or seventh cranial nerve; (4) The duration of cranial nerve palsy is in the range of approximately 15 to 262 days, and optionally, the median duration of cranial nerve palsy is approximately 77 days; or (5) Treatment may include corticosteroids; (d) CAR-T cell neurotoxicity is peripheral neuropathy, and peripheral neuropathy occurs in approximately 2.8% of subjects on an optional basis; or (e) CAR-T cell neurotoxicity is an adverse event that occurs during the study treatment for motor and neurocognitive function, and the adverse event that occurs during the study treatment for motor and neurocognitive function is of grade 1, and further randomly selected, the proportion of subjects experiencing a grade 1 adverse event during the study treatment for motor and neurocognitive function is approximately 0.6%. The method described in Embodiment 87. 89. The method according to any one of embodiments 67 to 88, wherein the subject has one or more high-risk cytogenetic abnormalities selected from the group including Gain / amp(1q), del(17p), t(4;14), t(14;16), or any combination thereof. 90. The method according to Embodiment 89, wherein the subject has at least two cytogenetic abnormalities, and optionally the subject has two, three, four, five, or more cytogenetic abnormalities. 91. The method according to Embodiment 89 or 90, wherein the cytogenetic abnormality is a standard-risk cytogenetic abnormality. 92. The method according to any one of Embodiments 1 to 91, further comprising administering a dose of T cells and then treating the subject for a CAR-T related adverse event, wherein the CAR-T related adverse event includes cytokine release syndrome (CRS) and / or neurotoxicity. 93. The method according to Embodiment 92, wherein the CRS CAR-T-related adverse event is a Grade 1 maximum toxicity grade CRS, and optionally, a Grade 1 maximum toxicity grade occurs in approximately 52.8% of the subjects. 94. The method according to Embodiment 92, wherein the CRS CAR-T-related adverse event is a Grade 2 maximum toxicity grade CRS, and optionally, a Grade 2 maximum toxicity grade occurs in approximately 22.2% of the subjects. 95. The method according to Embodiment 92, wherein the CRS CAR-T-related adverse event is a Grade 3 maximum toxicity grade CRS, and optionally, a Grade 3 maximum toxicity grade occurs in approximately 1.1% of the subjects. 96. The method according to any one of embodiments 93 to 95, wherein the time to the onset of CRS is in the range of approximately 1 to approximately 23 days, and optionally, the median time to the onset of CRS is approximately 8 days. 97. The method according to any one of embodiments 93 to 96, wherein the duration of CRS is in the range of approximately 1 to approximately 17 days, and optionally, the duration of CRS is approximately 3 days on a median basis. 98. The method according to any one of embodiments 93 to 97, wherein the treatment comprises tocilizumab, oxygen, corticosteroids, vasopressors, or any combination thereof. 99. The method according to Embodiment 92, wherein the neurotoxic CAR-T related adverse event is selected from the group consisting of immune effector cell-associated neurotoxicity syndrome or associated symptoms, motor / neurocognitive neurotoxicity, neurotoxic adverse events occurring under investigational treatment, non-immune effector cell-associated neurotoxicity syndrome or associated symptoms, or any combination thereof. 100. The method according to Embodiment 92, wherein the neurotoxicity is immunoeffector cell-associated neurotoxic syndrome or associated symptoms. 101. The method according to Embodiment 100, wherein the target immunoeffector cell-associated neurotoxic syndrome or associated symptoms occur in approximately 4.5% of cases. 102. The method according to Embodiment 100 or 101, wherein the maximum toxicity grade of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is Grade 1, and the maximum toxicity grade of Grade 1 in the subjects occurs at a rate of approximately 3.4%. 103. The method according to Embodiment 100 or 101, wherein the maximum toxicity grade of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is Grade 2, and the maximum toxicity grade of Grade 1 in the subjects occurs at a rate of approximately 1.1%. 104. The method according to any one of Embodiments 100 to 103, wherein the time to the onset of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is in the range of approximately 6 to approximately 15 days. 105. The method according to Embodiment 104, wherein the median time to the onset of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is approximately 9.5 days. 106. The method according to any one of Embodiments 100 to 103, wherein the duration of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is in the range of approximately 1 to approximately 6 days. 107. The method according to Embodiment 106, wherein the median duration of immunoeffector cell-associated neurotoxicity syndrome or associated symptoms is approximately 2 days. 108. The method according to any one of embodiments 100 to 107, wherein the treatment comprises a corticosteroid and / or tocilizumab. 109. The method according to any one of embodiments 99 to 108, wherein the target CAR-T cell neurotoxicity occurs at a rate of approximately 17.0%. 110. The method according to any one of Embodiments 99 to 109, wherein the CAR-T cell neurotoxicity is grade 3 / 4 neurotoxicity, and grade 3 / 4 neurotoxicity occurs in approximately 2.3% of subjects. 111. The method according to any one of embodiments 99 to 109, wherein the CAR-T cell neurotoxicity is grade 5 neurotoxicity. 112. The method according to any one of Embodiments 99 to 109, wherein the CAR-T cell neurotoxicity is cranial nerve palsy, and cranial nerve palsy occurs in approximately 9.1% of the subjects. 113. The method according to Embodiment 112, wherein the cranial nerve palsy is of grade 2, and grade 2 cranial nerve palsy occurs in approximately 8.0% of the subjects. 114. The method according to Embodiment 112, wherein the cranial nerve palsy is of grade 3, and grade 2 cranial nerve palsy occurs in approximately 1.1% of the subjects. 115. The method according to any one of embodiments 112 to 114, wherein the period from administration of a dose of T cells to the onset of cranial nerve palsy is in the range of approximately 17 days to approximately 60 days. 116. The method according to any one of embodiments 112 to 115, wherein the cranial nerve palsy occurs in the third, fifth, or seventh cranial nerve. 117. The method according to any one of embodiments 112 to 116, wherein the duration of cranial nerve palsy is in the range of approximately 15 days to approximately 262 days. 118. The method according to any one of embodiments 112 to 117, wherein the treatment comprises a corticosteroid. 119. The method according to any one of Embodiments 99 to 109, wherein the CAR-T cell neurotoxicity is peripheral neuropathy, and peripheral neuropathy occurs in approximately 2.8% of the subjects. 120. The method according to any one of Embodiments 99 to 109, wherein CAR-T cell neurotoxicity is the most frequent adverse event (MNT) observed during the experimental treatment for motor and neurocognitive function. 121. The method according to Embodiment 120, wherein the MNTs are Grade 1 MNTs, and Grade 1 MNTs occur at a rate of approximately 0.6% in the subject. 122. The method according to Embodiment 64 or 89, wherein the cytogenetic abnormality includes Gain / amp(1q). 123. The method according to Embodiment 64 or 89, wherein the cytogenetic abnormality includes del(17p). 124. The method according to Embodiment 64 or 89, wherein the cytogenetic abnormality includes t(4;14). 125. The method according to Embodiment 64 or 89, wherein the cytogenetic abnormality includes t(14;16). 126. The method according to any one of Embodiments 1 to 91, wherein the subject exhibits a reduced risk of developing CAR-T related adverse events. 127. The method according to Embodiment 126, wherein the CAR-T related adverse event includes cytokine release syndrome (CRS). 128. The method according to Embodiment 127, wherein CRS occurs in the subject at a rate of approximately 60% to approximately 90%, or approximately 76.1%. 129. The method according to Embodiment 127 or 128, wherein the maximum toxicity grade of CRS is Grade 1, Grade 2, or Grade 3. 130. The method according to any one of Embodiments 126 to 129, wherein the maximum toxicity grade of CRS is Grade 1, and optionally, the maximum toxicity grade of Grade 1 in the subject occurs at a rate of approximately 52.8%. 131. The method according to any one of Embodiments 126 to 130, wherein the maximum toxicity grade of CRS is Grade 2, and optionally, the maximum toxicity grade of Grade 2 occurs in approximately 22.2% of the subjects. 132. The method according to any one of embodiments 126 to 131, wherein the maximum toxicity grade of CRS is Grade 3, and optionally, the maximum toxicity grade of Grade 3 in the subject occurs at a rate of approximately 1.1%. 133. The method according to Embodiment 126, wherein CAR-T related adverse events include neurotoxicity. 134. The method according to Embodiment 133, wherein the neurotoxic CAR-T related adverse event is selected from the group consisting of immune effector cell-associated neurotoxicity syndrome or associated symptoms, motor / neurocognitive neurotoxicity, neurotoxic adverse events occurring under investigational treatment, non-immune effector cell-associated neurotoxicity syndrome or associated symptoms, or any combination thereof. 135. The method according to Embodiment 134, wherein the neurotoxicity is immunoeffector cell-associated neurotoxic syndrome or associated symptoms. 136. The method according to Embodiment 135, wherein the target immunoeffector cell-associated neurotoxic syndrome or associated symptoms occur in approximately 4.5% of cases. 137. The method according to Embodiment 135 or 136, wherein the maximum toxicity grade of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is Grade 1, and the maximum toxicity grade of Grade 1 in the subjects occurs at a rate of approximately 3.4%. 138. The method according to Embodiment 135 or 136, wherein the maximum toxicity grade of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is Grade 2, and the maximum toxicity grade of Grade 1 in the subjects occurs at a rate of approximately 1.1%. 139. The method according to any one of embodiments 135 to 138, wherein the time to the onset of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is in the range of approximately 6 to approximately 15 days. 140. The method according to Embodiment 139, wherein the median time to the onset of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is approximately 9.5 days. 141. The method according to any one of embodiments 135 to 138, wherein the duration of immunoeffector cell-associated neurotoxic syndrome or associated symptoms is in the range of approximately 1 to approximately 6 days. 142. The method according to Embodiment 141, wherein the median duration of immunoeffector cell-associated neurotoxicity syndrome or associated symptoms is approximately 2 days. 143. The method according to any one of Embodiments 135 to 142, wherein the treatment comprises a corticosteroid and / or tocilizumab. 144. The method according to any one of Embodiments 134 to 143, wherein the target CAR-T cell neurotoxicity occurs at a rate of approximately 17.0%. 145. The method according to any one of Embodiments 134 to 144, wherein the CAR-T cell neurotoxicity is grade 3 / 4 neurotoxicity, and grade 3 / 4 neurotoxicity occurs in approximately 2.3% of subjects. 146. The method according to any one of Embodiments 134 to 145, wherein the CAR-T cell neurotoxicity is Grade 5 neurotoxicity. 147. The method according to any one of Embodiments 134 to 144, wherein the CAR-T cell neurotoxicity is cranial nerve palsy, and cranial nerve palsy occurs in approximately 9.1% of the subjects. 148. The method according to Embodiment 147, wherein the cranial nerve palsy is of grade 2, and grade 2 cranial nerve palsy occurs in approximately 8.0% of the subjects. 149. The method according to Embodiment 147, wherein the cranial nerve palsy is of grade 3, and grade 2 cranial nerve palsy occurs in approximately 1.1% of the subjects. 150. The method according to any one of embodiments 147 to 149, wherein the period from administration of a dose of T cells to the onset of cranial nerve palsy is in the range of approximately 17 days to approximately 60 days. 151. The method according to any one of embodiments 147 to 150, wherein the cranial nerve palsy occurs in the third, fifth, or seventh cranial nerve. 152. The method according to any one of embodiments 147 to 151, wherein the duration of cranial nerve palsy is in the range of approximately 15 days to approximately 262 days. 153. The method according to any one of embodiments 147 to 152, wherein the treatment comprises a corticosteroid. 154. The method according to any one of Embodiments 134 to 144, wherein the CAR-T cell neurotoxicity is peripheral neuropathy, and peripheral neuropathy occurs in approximately 2.8% of the subjects. 155. The method according to any one of Embodiments 134 to 144, wherein CAR-T cell neurotoxicity is the most frequent adverse event (MNT) observed during the experimental treatment for motor and neurocognitive function. 156. The method according to Embodiment 155, wherein the MNTs are Grade 1 MNTs, and Grade 1 MNTs occur at a rate of approximately 0.6% in the subject. 157. Peripheral neuropathy is grade 1, and grade 1 peripheral neuropathy occurs in approximately 1.1% of cases in embodiments 38, 63, 88, 119, and 154. 158. The method according to any one of Embodiments 38, 63, 88, 119, and 154, wherein peripheral neuropathy is grade 2, and grade 2 peripheral neuropathy occurs at a rate of approximately 1.1%. 159. The method according to any one of Embodiments 38, 63, 88, 119, and 154, wherein peripheral neuropathy is grade 3, and grade 3 peripheral neuropathy occurs at a rate of approximately 0.6%. 160. A method for selectively treating subjects, wherein subjects determined to have high-risk characteristics, (a) Extracellular antigen-binding domain having specific binding ability to BCMA epitopes, (b) Transmembrane domain, and (c) Intracellular signaling domain A method comprising administering a certain dose of chimeric antigen receptor (CAR)-containing T cells, CAR contains the amino acid sequence of SEQ ID NO: 17, High-risk features include cytogenetic abnormalities, International Staging System (ISS) stage III, and / or soft tissue plasmacytoma. The subjects were those with multiple myeloma who had received 1 to 3 prior treatment lines, including IMiD therapy, and who were resistant to IMiD. The method is effective in achieving complete efficacy as described in Example 3, any one of Tables 1 to 5 and Table 15, or any one of Figures 6A to 6C, Figure 7, and Figures 8A to 8D. Optionally, the method further includes treating the subject for adverse events after administering a dose of T cells as described in Example 4, or any one of Tables 6 to 14 and Table 16, and A method further comprising, optionally, administering a treatment to a subject to mitigate an adverse event as described in Example 4, or in any one of Tables 6 to 14 and Table 16. [Examples]

[0251] The following examples are provided to further illustrate some of the embodiments and models disclosed herein. These examples are intended to illustrate, rather than limit, the embodiments or models disclosed.

[0252] Example 1: Siltacaptagene autoleucel B cell maturation antigen (BCMA, also known as CD269 and TNFRSF17) is a 20-kilodalton, type III membrane protein that is part of the tumor necrosis receptor superfamily. BCMA is a cell surface antigen that is predominantly expressed at high levels in B lineage cells. Figure 1 shows BCMA expression on various immune-derived cells. Comparative studies have shown BCMA deficiency in most normal tissues and lack of expression on CD34-positive hematopoietic stem cells. BCMA binds to two ligands that induce B cell proliferation and plays a crucial role in B cell maturation and subsequent differentiation into plasma cells. Its selective expression and biological importance for myeloma cell proliferation and survival make BCMA a promising target for CAR-T-based immunotherapy.

[0253] Silta-cel is an autologous chimeric antigen receptor T cell (CAR-T) therapy that targets BCMA. The silta-cel chimeric antigen receptor (CAR) contains two BCMA-targeting VHH domains designed to confer avidity. A construct map is shown in Figure 2, and a schematic diagram of CAR-T cell generation is shown in Figure 3. Cilta-cel contains a VHH domain with the amino acid sequence shown in SEQ ID NO: 2 and a VHH domain with the amino acid sequence shown in SEQ ID NO: 4.

[0254] Example 2: Treatment method with siltacaptagen autoleucel Cilta-cel is highly effective in relapsed / resistant multiple myeloma (RRMM) with a severe history of prior treatment. In this study, the inventors investigated cilta-cel in past treatment lines in lenalidomide-resistant patients.

[0255] Many multiple myeloma (MM) patients experience relapse after standard treatment (van de Donk, Hematology Am Soc Hematol Educ Program 2020;2020:248-58; Rodriguez-Lobato et al., Br J Haematol 2022;196:649-59), and their outcomes worsen with each subsequent line of treatment (LOT) (Yong et al., Br J Haematol 2016;175:252-64; Dhakal et al., Clinical Lymphoma Myeloma and Leukemia 2022;22:S167; Dhakal et al., HemaSphere 2022;6:790-1). Lenalidomide is a recommended immunomodulatory agent for newly diagnosed multifilariatic tumors (MM) and relapsed / resistant MM (RRMM) (Dimopoulos et al., HemaSphere 2021;5:e528; National Comprehensive Cancer Network. NCCN clinical practice guidelines in oncology, (NCCN Guidelines®) version 3.2023.2023). The use of lenalidomide is widespread in initial line settings, including maintenance therapy (van de Donk, Hematology Am Soc Hematol Educ Program 2020;2020:248-58; de Arriba de la Fuente et al., Cancers(Basel)2022;15). The increasing rate of lenalidomide resistance in the early stages of treatment (van de Donk, Hematology Am Soc Hematol Educ Program 2020;2020:248-58; de Arriba de la Fuente et al., Cancers (Basel) 2022;15) highlights the growing need for new and effective therapies for lenalidomide-resistant diseases (de Arriba de la Fuente et al., Cancers (Basel) 2022;15).The high treatment discontinuation rate—only 13% to 35% of patients receive 4 or more lots—highlights the need for early and effective treatment (Fonseca et al., BMC Cancer 2020;20:1087).

[0256] Cilta-cel led to early, deep, and sustained responses in patients with RRMM and three or more prior LOTs in the Phase 1b / 2 CARTITUDE-1 trial (median progression-free survival [PFS] of 34.9 months) (Berdeja et al., Lancet 2021;398:314-24; Martin et al., J Clin Oncol 2022:JCO2200842; Lin et al., J Clin Oncol 2023. Submitted). The Phase 2 CARTITUDE-2 study (Cohorts A and B) demonstrated the efficacy of cilta-cel with a response rate of 95%–100% in a small cohort of early-stage disease patients, and the median duration of response (DOR) and median progression-free survival (PFS) were not reached even after approximately 1.5 years of follow-up (van de Donk et al., Blood 2022;140:7536-7; Einsele et al., American Society of Clinical Oncology Annual Meeting; 2022, June 3-7; Chicago, IL).

[0257] CARTITUDE-4 is a phase 3 randomized controlled trial comparing cilta-cel to physician's choice between two highly effective standard treatments in lenalidomide-resistant MM patients after 1–3 lots. We report the efficacy and safety results from the initially planned analysis of CARTITUDE-4.

[0258] Study design and patients CARTITUDE-4 is a global, open-label, randomized phase 3 trial conducted at 81 sites in the United States, Europe, Asia, and Australia. Eligible patients were lenalidomide-resistant (Rajkumar et al., Blood 2011;117:4691-5), had 1 to 3 prior LOTs including proteasome inhibitors and immunomodulators, had an Eastern Cooperative Oncology Group Performance Status score of ≤1, had no prior CAR-T therapy history, and had not previously received BCMA-targeted therapy.

[0259] Randomization and treatment Patients were randomly assigned in a 1:1 ratio via computer-generated randomization to receive either physician-selected standard treatment (pomalidomide-bortezomib-dexamethasone [PVd] (Richardson et al., Lancet Oncol 2019;20:781-94) or daratumumab-pomalidomide-dexamethasone [DPd] (Dimo...

Claims

1. A method for treating the subject, wherein the subject (a) Extracellular antigen-binding domain having the ability to specifically bind to the epitope of B cell maturation antigen (BCMA), (b) Transmembrane domain, and (c) Intracellular signal transduction domain A method comprising administering a certain dose of chimeric antigen receptor (CAR)-containing T cells, A method wherein the subject has multiple myeloma, has received one to three prior treatment lines, including immunomodulatory drug (IMiD) therapy, and is resistant to said IMiD.

2. The method according to claim 1, wherein the subject has high-risk features, and optionally the high-risk features are a cytogenetic abnormality, International Staging System (ISS) stage III, and / or soft tissue plasmacytoma.

3. A method of selectively treating a target, (1) Determining whether the subject has high-risk characteristics, wherein the high-risk characteristics are cytogenetic abnormalities, International Staging System (ISS) stage III, and / or soft tissue plasmacytoma; and (2) The subject determined to have the high-risk characteristics in step (1) (a) Extracellular antigen-binding domain having specific binding ability to BCMA epitopes, (b) Transmembrane domain, and (c) Intracellular signal transduction domain Administering a certain dose of chimeric antigen receptor (CAR)-containing T cells, In a method including, A method in which, on an optional basis, the subject has multiple myeloma, has received one to three prior treatment lines, including therapy with IMiD, and is resistant to IMiD.

4. A method for selectively treating a subject, wherein the subject is determined to have high-risk characteristics, (a) Extracellular antigen-binding domain having specific binding ability to BCMA epitopes, (b) Transmembrane domain, and (c) Intracellular signal transduction domain A method comprising administering a certain dose of chimeric antigen receptor (CAR)-containing T cells, The aforementioned high-risk features are cytogenetic abnormalities, International Staging System (ISS) stage III, and / or soft tissue plasmacytoma. A method in which, on an optional basis, the subject has multiple myeloma, has received one to three prior treatment lines, including therapy with IMiD, and is resistant to IMiD.

5. The method according to any one of claims 1 to 4, wherein the IMiD is lenalidomide.

6. The method according to any one of claims 2 to 5, wherein the high-risk feature is a cytogenetic abnormality.

7. The method according to claim 6, wherein the cytogenetic abnormality is a high-risk cytogenetic abnormality.

8. The method according to claim 7, wherein the subject has one or more high-risk cytogenetic abnormalities selected from the group including Gain / amp(1q), del(17p), t(4;14), t(14;16), or any combination thereof.

9. The method according to claim 8, wherein the cytogenetic abnormality includes Gain / amp(1q).

10. The method according to claim 8, wherein the cytogenetic abnormality includes del(17p).

11. The method according to claim 8, wherein the cytogenetic abnormality includes t(4;14).

12. The method according to claim 8, wherein the cytogenetic abnormality includes t(14;16).

13. The method according to any one of claims 6 to 12, wherein the subject has at least two cytogenetic abnormalities, and optionally the subject has two, three, four, five, or more cytogenetic abnormalities.

14. The method according to claim 6, wherein the cytogenetic abnormality is a standard-risk cytogenetic abnormality.

15. The method according to any one of claims 2 to 5, wherein the high-risk characteristic is International Staging System (ISS) stage III.

16. The method according to any one of claims 2 to 5, wherein the high-risk feature is a soft tissue plasmacytoma.

17. The method according to any one of claims 1 to 16, wherein the subject has received one prior treatment line.

18. The method according to any one of claims 1 to 16, wherein the subject has received two prior treatment lines.

19. The method according to any one of claims 1 to 16, wherein the subject has received three prior treatment lines.

20. The method according to any one of claims 1 to 19, wherein the 1, 2, or 3 prior treatment lines include treatment with pomalidomide.

21. The method according to any one of claims 1 to 20, wherein the 1, 2, or 3 prior treatment lines further comprise treatment with an anti-CD38 antibody, and optionally the anti-CD38 antibody is daratumumab and / or isatuximab.

22. The method according to any one of claims 1 to 21, wherein the 1, 2, or 3 prior treatment lines further comprise treatment with a proteasome inhibitor, wherein the proteasome inhibitor is bortezomib, carfilzomib, ixazomib, or any combination thereof.

23. The method according to any one of claims 1 to 22, wherein the subject has previously received bridging therapy, the bridging therapy is at the discretion of the physician, the bridging therapy comprises pomalidomide, bortezomib, dexamethasone, daratumumab, or any combination thereof, the bridging therapy comprises pomalidomide, bortezomib and dexamethasone, and the bridging therapy comprises daratumumab, pomalidomide and dexamethasone.

24. The method according to claim 23, wherein the subject has received the bridging therapy approximately every 20 to 30 days, optionally, the subject has received the bridging therapy approximately every 21 days, optionally, the subject has received the bridging therapy approximately every 28 days, and optionally, the subject has received at least one, two, three, four, or more bridging therapies.

25. The aforementioned subjects have previously received lymphocyte depletion therapy, and optionally, the lymphocyte depletion therapy includes daily cyclophosphamide and / or fludarabine, and optionally, the lymphocyte depletion therapy includes daily cyclophosphamide and fludarabine, and optionally, the lymphocyte depletion therapy includes approximately 300 mg / m² daily for three days. ２ The concentration of cyclophosphamide is approximately 30 mg / m³. ２ The method according to any one of claims 1 to 24, comprising fludarabine at the concentration of [amount].

26. The dose of the T cells is 0.5 to 1.0 × 10 ６ Cells / Body weight of the subject is 1 kg, and optionally, the dose of T cells is approximately 0.75 × 10 ６ The method according to any one of claims 1 to 25, wherein the cells are 1 kg of the subject's body weight, and optionally the method comprises administering the dose of T cells about 5 to 7 days after the start of the lymphocyte depletion therapy, and optionally the dose is administered as a single infusion.

27. The method according to any one of claims 1 to 26, which is effective in achieving a complete response in the subject after administering the T cells in the aforementioned dose to the subject, which is optionally effective in achieving the complete response in a ratio of approximately 75% to approximately 100%, which is optionally effective in achieving the complete response in a ratio of approximately 84.6%, and which is optionally effective in achieving the complete response in a ratio of approximately 99.4%.

28. The aforementioned overall response rates are listed in order from best to worst. (1) Strict complete response; (2) Complete response; (3) Best partial response; (4) Partial response; or (5) Minimum response The method according to claim 27, including the method described in claim 27.

29. The method according to claim 28, wherein the overall response is a strict complete response, and the method is effective in achieving the strict complete response in a rate of approximately 40% to approximately 90%, approximately 50% to approximately 80%, approximately 58.2%, or approximately 68.8%, at any discretion.

30. The method according to claim 28, wherein the overall response is a complete response, and the method is optionally effective in achieving the complete response in a ratio of approximately 10% to approximately 20%, and further optionally effective in achieving the complete response in a ratio of approximately 14.9% or approximately 17.6%.

31. The method according to claim 28, wherein the overall response is the best partial response or a partial response.

32. (1) Effective in achieving strict complete response or complete response in approximately 70% to 90% of cases, and, on an optional basis, effective in achieving strict complete response or complete response in approximately 73.1% or 86.4% of cases; (2) Effective in achieving strict complete response, complete response, or best partial response in approximately 80% to 100% of cases, and optionally effective in achieving strict complete response, complete response, or best partial response in approximately 81.3% or 96.0% of cases; (3) Effective in achieving minimal response; (4) Effective in further achieving minimal residual disease negativity, and optionally effective in achieving minimal residual disease negativity in approximately 50% to 80% of cases, and optionally effective in achieving minimal residual disease negativity in approximately 60.6% or 71.6% of cases; or (5) It is effective in helping at least approximately 60-100% of subjects, at least approximately 69.4-81.1% of subjects, or at least approximately 84.1-93.4% of subjects to achieve further 12-month progression-free survival, and on a voluntary basis, it is effective in helping at least approximately 75.9% of subjects or approximately 89.7% of subjects to achieve 12-month progression-free survival. The method according to claim 28.

33. (1) The time to complete response or minimum response on the first attempt is in the range of approximately 0.9 to approximately 11.1 months, and optionally, the median time to complete response or minimum response on the first attempt is approximately 2.1 months; or (2) The time to best overall response or best minimum response is approximately 1.1 to approximately 18.6 months, and optionally, the median time to the best overall response or best minimum response is approximately 6.4 or approximately 6.5 months. The method according to any one of claims 27 to 32.

34. The method further comprises treating the subject with respect to an adverse event after administering the dose of the T cells, optionally comprising administering a treatment to the subject to mitigate the adverse event, optionally comprising the adverse event being a hematological adverse event, a non-hematological adverse event, an adverse event occurring under the study treatment, or any combination thereof, optionally comprising the non-hematological adverse event being an infection and / or a non-hematological adverse event other than an infection, and optionally comprising the adverse event being neutropenia, thrombocytopenia, anemia, lymphopenia, upper respiratory tract infection, nasopharyngitis, sinusitis, rhinitis, tonsillitis, pharyngitis, laryngitis, pharyngotonsillitis, COVID-19, COVID-19 pneumonia, asymptomatic COVID-19, neutropenic sepsis, progressive multifocal whitehead The method according to any one of claims 1 to 33, comprising progressive multifocal leukoencephalopathy, septic shock, respiratory failure, pulmonary embolism, lower respiratory tract / lung infection, pneumonia, bronchitis, nausea, hypogammaglobulinemia, diarrhea, fatigue, headache, constipation, hypokalemia, asthenia, peripheral edema, decreased appetite, peripheral sensory neuropathy, back pain, arthralgia, fever, dyspnea, insomnia, or any combination thereof, further optionally, the adverse event is a grade 3 / 4 adverse event, and further optionally, the adverse event persists for more than about 30 days or about 60 days.

35. The method according to any one of claims 1 to 34, wherein the method further comprises treating the subject with respect to a secondary primary malignancy after administering the T cells in the dose said, optionally comprising administering a treatment to the subject to alleviate the secondary primary malignancy, optionally comprising the secondary primary malignancy comprising cutaneous / non-invasive malignancy, hematological malignancy, non-cutaneous / invasive malignancy, or any combination thereof, and further optionally comprising basal cell carcinoma, Bowen's disease, squamous cell carcinoma of the lip, malignant melanoma, malignant melanoma in situ, squamous cell carcinoma of the skin, acute myeloid leukemia, myelodysplastic syndrome, peripheral T-cell lymphoma, angiosarcoma, invasive lobular breast carcinoma, pleomorphic malignant fibrous histiocytoma, renal cell carcinoma, tonsil carcinoma, or any combination thereof.

36. The method according to claim 34 or 35, wherein the adverse event or secondary primary malignant tumor occurs in the subject at a rate equivalent to the rate at which the same adverse event or the same secondary primary malignant tumor occurs in a subject receiving standard treatment.

37. The method further comprises administering the T cells in the dose described above and then treating the subject for CAR-T related adverse events, optionally comprising administering a treatment to the subject to mitigate the CAR-T related adverse events, optionally comprising the CAR-T related adverse events including cytokine release syndrome (CRS) and / or neurotoxicity, optionally, (a) The CAR-T-related adverse event is CRS, and optionally, the CRS occurs in approximately 60% to 90% or 76.1% of the subjects, and optionally, the maximum toxicity grade of the CRS is Grade 1, Grade 2 or Grade 3, and optionally, (1) The maximum toxicity grade of the CRS is Grade 1, and optionally, the maximum toxicity grade of Grade 1 occurs in approximately 52.8% of the subjects; (2) The maximum toxicity grade of the CRS is Grade 2, and optionally, the maximum toxicity grade of Grade 2 occurs in approximately 22.2% of the subjects; (3) The maximum toxicity grade of the CRS is Grade 3, and optionally, the maximum toxicity grade of Grade 3 occurs in approximately 1.1% of the subjects; (4) The period until the first onset of CRS is in the range of approximately 1 to approximately 23 days, and optionally, the median period until the first onset of CRS is approximately 8 days; (5) The duration of the CRS is in the range of approximately 1 to approximately 17 days, and optionally, the median duration of the CRS is approximately 3 days; or (6) The treatment includes tocilizumab, oxygen, corticosteroids, vasopressors, or any combination thereof; or (b) The CAR-T related adverse event is neurotoxic, and optionally, the neurotoxicity includes immunoeffector cell-associated neurotoxic syndrome or associated symptoms, motor / neurocognitive neurotoxicity, neurotoxic adverse events occurring under study treatment, non-immune effector cell-associated neurotoxic syndrome or associated symptoms, or any combination thereof, and optionally, the neurotoxicity is immunoeffector cell-associated neurotoxic syndrome or associated symptoms, and optionally, (1) The immunoeffector cell-associated neurotoxic syndrome or associated symptoms occur in approximately 4.5% of the subjects; (2) The maximum toxicity grade of the immunoeffector cell-associated neurotoxic syndrome or associated symptoms is Grade 1 or Grade 2, and optionally, the maximum toxicity grade of the immunoeffector cell-associated neurotoxic syndrome or associated symptoms is Grade 1, and optionally, the maximum toxicity grade of Grade 1 occurs in approximately 3.4% of the subjects, or optionally, the maximum toxicity grade of the immunoeffector cell-associated neurotoxic syndrome or associated symptoms is Grade 2, and optionally, the maximum toxicity grade of Grade 2 occurs in approximately 1.1% of the subjects; (3) The period until the onset of the immunoeffector cell-associated neurotoxic syndrome or associated symptoms is in the range of approximately 6 to approximately 15 days, and optionally, the median period until the onset of the immunoeffector cell-associated neurotoxic syndrome or associated symptoms is approximately 9.5 days; (4) The duration of the immunoeffector cell-associated neurotoxic syndrome or associated symptoms is in the range of approximately 1 to approximately 6 days, and optionally, the median duration of the immunoeffector cell-associated neurotoxic syndrome or associated symptoms is approximately 2 days; or (5) The treatment includes a corticosteroid and / or tocilizumab, The method according to any one of claims 1 to 36.

38. The neurotoxicity is CAR-T cell neurotoxicity, and optionally, the CAR-T cell neurotoxicity occurs in approximately 17.0% of the subjects, and optionally, the CAR-T cell neurotoxicity includes grade 3 / 4 neurotoxicity, grade 5 neurotoxicity, cranial nerve palsy, peripheral neuropathy, adverse events occurring during the study treatment relating to motor / neurocognitive function, or any combination thereof, and optionally, (a) The CAR-T cell neurotoxicity is a grade 3 / 4 neurotoxicity occurring in approximately 2.3% of the subjects; (b) The CAR-T cell neurotoxicity is Grade 5 neurotoxicity; (c) The CAR-T cell neurotoxicity is cranial nerve palsy, and in the subjects, the cranial nerve palsy occurs at a rate of approximately 9.1%, and in the subjects, (1) The cranial nerve palsy is of grade 2 or grade 3, and optionally, the cranial nerve palsy is of grade 2, occurring in approximately 8.0% of the subjects, and optionally, the cranial nerve palsy is of grade 3, occurring in approximately 1.1% of the subjects; (2) The period from administration of the T cells in the dose described above to the onset of cranial nerve palsy is in the range of approximately 17 to approximately 60 days, and optionally, the median period from administration of the T cells in the dose described above to the onset of cranial nerve palsy is approximately 21 days; (3) The cranial nerve palsy occurs in the third, fifth, or fifth cranial nerve; (4) The duration of the cranial nerve palsy is in the range of approximately 15 days to approximately 262 days, and optionally, the median duration of the cranial nerve palsy is approximately 77 days; or (5) The treatment includes corticosteroids; (d) The CAR-T cell neurotoxicity is peripheral neuropathy, and optionally, peripheral neuropathy occurs in approximately 2.8% of the subjects; or (e) The CAR-T cell neurotoxicity is an adverse event that occurred during the experimental treatment for motor and neurocognitive function, and the adverse event that occurred during the experimental treatment for motor and neurocognitive function is of grade 1, and further, the adverse event of grade 1 that occurred during the experimental treatment for motor and neurocognitive function occurs in approximately 0.6% of the subjects, The method according to claim 37.

39. (a) The number of CD3+ cells containing the CAR in the subject's blood reaches a peak approximately 13 days after administration of the T cells to the subject, and optionally, the number of CD3+ cells containing the CAR in the subject's blood reaches a peak at an average concentration of approximately 1523 cells / μL; (b) CD3+ cells containing the CAR in the subject's blood remain detectable approximately 13 to 631 days after the administration of the T cells to the subject, and optionally, CD3+ cells containing the CAR in the subject's blood remain detectable approximately 57 days after the administration of the T cells to the subject; or (c) AUC of CD3+ cells containing the CAR in the blood of the subject. ０－２８ However, the average value is approximately 12,504 cells / μL. The method according to any one of claims 1 to 38.

40. The first VHH domain includes CDR1, CDR2, and CDR3 as shown in the VHH domain containing the amino acid sequence of SEQ ID NO: 2, and the second VHH domain includes CDR1, CDR2, and CDR3 as shown in the VHH domain containing the amino acid sequence of SEQ ID NO: 4, optionally the first VHH domain includes CDR1 containing the amino acid sequence of SEQ ID NO: 18, CDR2 containing the amino acid sequence of SEQ ID NO: 19, and CDR3 containing the amino acid sequence of SEQ ID NO: 20, and the second VHH The H domain includes CDR1 containing the amino acid sequence of SEQ ID NO: 21, CDR2 containing the amino acid sequence of SEQ ID NO: 22, and CDR3 containing the amino acid sequence of SEQ ID NO: 23, and optionally the first VHH domain contains the amino acid sequence of SEQ ID NO: 2, and the second VHH domain contains the amino acid sequence of SEQ ID NO: 4, and optionally the first VHH domain is located on the N-terminal side of the second VHH domain, or the first VHH domain is located on the C-terminal side of the second VHH domain, and optionally, (a) The first VHH domain is linked to the second VHH domain by a linker containing the amino acid sequence of SEQ ID NO: 3; (b) The transmembrane domain is derived from a molecule selected from the group consisting of CD8α, CD4, CD28, CD137, CD80, CD86, CD152 and PD1, and optionally the transmembrane domain is derived from CD8α and contains the amino acid sequence of SEQ ID NO: 6; (c) The intracellular signaling domain includes a primary intracellular signaling domain of an immune effector cell, and optionally, the primary intracellular signaling domain is derived from CD3ζ containing the amino acid sequence of SEQ ID NO: 8; (d) The intracellular signaling domain comprises a co-stimulatory signaling domain, optionally, the co-stimulatory signaling domain is derived from a co-stimulatory molecule selected from the group consisting of ligands CD27, CD28, CD137, OX40, CD30, CD40, CD3, LFA-1, ICOS, CD2, CD7, LIGHT, NKG2C, B7-H3, CD83 and any combination thereof, optionally, the co-stimulatory signaling domain comprises the cytoplasmic domain of CD137 containing the amino acid sequence of SEQ ID NO: 7; (e) The CAR further comprises a hinge domain located between the C-terminus of the extracellular antigen-binding domain and the N-terminus of the transmembrane domain, wherein the hinge domain is optionally derived from CD8α having the amino acid sequence of SEQ ID NO: 5; (f) The CAR further comprises a signal peptide located at the N-terminal end of the polypeptide, wherein the signal peptide is derived from CD8α having the amino acid sequence of SEQ ID NO: 1; or (g) The CAR contains the amino acid sequence of SEQ ID NO: 17, The method according to any one of claims 1 to 39.

41. The method according to any one of claims 1 to 40, wherein the T cells in the aforementioned dose are formulated into a composition containing 5% dimethyl sulfoxide (DMSO).

42. The method according to any one of claims 1 to 41, wherein administering the aforementioned dose of T cells reduces the risk of disease progression or death in the subject.

43. The method according to claim 42, wherein the risk of disease progression or death is reduced compared to the administration of daratumumab-pomalidomide-dexamethasone (DPd) or pomalidomide-bortezomib-dexamethasone (PVd) treatment.

44. The method according to claim 42, wherein the risk of disease progression or death is reduced compared to the administration of ide-cel therapy.

45. The method according to any one of claims 42 to 44, wherein the subject exhibits a disease progression or mortality risk reduced by approximately 60% to approximately 75%.

46. The method according to claim 45, wherein the subject exhibits a disease progression or mortality risk reduced by approximately 74%.

47. The method according to any one of claims 42 to 46, wherein the IMiD is lenalidomide.

48. The method according to any one of claims 42 to 47, wherein the subject has received three or fewer prior treatment lines.

49. The method according to any one of claims 42 to 47, wherein the subject has received two or fewer prior treatment lines.

50. The method according to any one of claims 42 to 47, wherein the subject has received only one prior treatment line.

51. The method according to any one of claims 42 to 50, wherein the method is effective in achieving an overall response rate (ORR) of approximately 75% to approximately 100%.

52. The method according to claim 51, wherein the ORR is approximately 84.6%.

53. The method according to any one of claims 42 to 52, wherein the treatment is effective in extending the median progression-free survival (PFS) of the subject compared to the administration of DPd or PVd treatment.

54. The method according to claim 53, wherein the PFS at 12 months after administration of the treatment is approximately 75.9%.

55. The method according to claim 53 or 54, wherein the PFS at 12 months after administration of DPd or PVd is approximately 48.6%.

56. The method according to any one of claims 1 to 23 and 42 to 55, wherein the treatment is more effective in achieving a strict complete response (sCR) in the subject compared to the administration of DPd or PVd treatment.

57. The method according to claim 56, wherein the sCR after administration of the treatment is approximately 58.2%.

58. The method according to claim 56 or 57, wherein the sCR after administration of DPd or PVd is approximately 15.2%.

59. The method according to any one of claims 1 to 23 and 42 to 58, wherein the treatment is more effective in achieving best partial response (VGPR) or better in the subject compared to the administration of DPd or PVd treatment.

60. The method according to claim 59, wherein approximately 81.3% of patients achieve a VGPR level of 2 or higher after administration of the aforementioned treatment.

61. The method according to claim 59 or 60, wherein approximately 45.5% of patients achieve a VGPR of 1 or higher after administration of DPd or PVd.

62. The method further comprises administering the T cells in the dose described above and then treating the subject for CAR-T related adverse events, optionally comprising administering a treatment to the subject to mitigate the CAR-T related adverse events, optionally comprising the CAR-T related adverse events including cytokine release syndrome (CRS) and / or neurotoxicity, optionally, (a) The CAR-T-related adverse event is CRS, and optionally, the CRS occurs in approximately 60% to 90% or 76.1% of the subjects, and optionally, the maximum toxicity grade of the CRS is Grade 1, Grade 2 or Grade 3, and optionally, (1) The maximum toxicity grade of the CRS is Grade 1, and optionally, the maximum toxicity grade of Grade 1 occurs in approximately 52.8% of the subjects; (2) The maximum toxicity grade of the CRS is Grade 2, and optionally, the maximum toxicity grade of Grade 2 occurs in approximately 22.2% of the subjects; (3) The maximum toxicity grade of the CRS is Grade 3, and optionally, the maximum toxicity grade of Grade 3 occurs in approximately 1.1% of the subjects; (4) The period until the first onset of CRS is in the range of approximately 1 to approximately 23 days, and optionally, the median period until the first onset of CRS is approximately 8 days; (5) The duration of the CRS is in the range of approximately 1 to approximately 17 days, and optionally, the median duration of the CRS is approximately 3 days; or (6) The treatment includes tocilizumab, oxygen, corticosteroids, vasopressors, or any combination thereof; or (b) The CAR-T related adverse event is neurotoxic, and optionally, the neurotoxicity includes immunoeffector cell-associated neurotoxic syndrome or associated symptoms, motor / neurocognitive neurotoxicity, neurotoxic adverse events occurring under study treatment, non-immune effector cell-associated neurotoxic syndrome or associated symptoms, or any combination thereof, and optionally, the neurotoxicity is immunoeffector cell-associated neurotoxic syndrome or associated symptoms, and optionally, (1) The immunoeffector cell-associated neurotoxic syndrome or associated symptoms occur in approximately 4.5% of the subjects; (2) The maximum toxicity grade of the immunoeffector cell-associated neurotoxic syndrome or associated symptoms is Grade 1 or Grade 2, and optionally, the maximum toxicity grade of the immunoeffector cell-associated neurotoxic syndrome or associated symptoms is Grade 1, and optionally, the maximum toxicity grade of Grade 1 occurs in approximately 3.4% of the subjects, or optionally, the maximum toxicity grade of the immunoeffector cell-associated neurotoxic syndrome or associated symptoms is Grade 2, and optionally, the maximum toxicity grade of Grade 2 occurs in approximately 1.1% of the subjects; (3) The period until the onset of the immunoeffector cell-associated neurotoxic syndrome or associated symptoms is in the range of approximately 6 to approximately 15 days, and optionally, the median period until the onset of the immunoeffector cell-associated neurotoxic syndrome or associated symptoms is approximately 9.5 days; (4) The duration of the immunoeffector cell-associated neurotoxic syndrome or associated symptoms is in the range of approximately 1 to approximately 6 days, and optionally, the median duration of the immunoeffector cell-associated neurotoxic syndrome or associated symptoms is approximately 2 days; or (5) The treatment includes a corticosteroid and / or tocilizumab, The method according to any one of claims 42 to 61.

63. The neurotoxicity is CAR-T cell neurotoxicity, and optionally, the CAR-T cell neurotoxicity occurs in approximately 17.0% of the subjects, and optionally, the CAR-T cell neurotoxicity includes grade 3 / 4 neurotoxicity, grade 5 neurotoxicity, cranial nerve palsy, peripheral neuropathy, adverse events occurring during the study treatment relating to motor / neurocognitive function, or any combination thereof, and optionally, (a) The CAR-T cell neurotoxicity is a grade 3 / 4 neurotoxicity occurring in approximately 2.3% of the subjects; (b) The CAR-T cell neurotoxicity is Grade 5 neurotoxicity; (c) The CAR-T cell neurotoxicity is cranial nerve palsy, and in the subjects, the cranial nerve palsy occurs at a rate of approximately 9.1%, and in the subjects, (1) The cranial nerve palsy is of grade 2 or grade 3, and optionally, the cranial nerve palsy is of grade 2, occurring in approximately 8.0% of the subjects, and optionally, the cranial nerve palsy is of grade 3, occurring in approximately 1.1% of the subjects; (2) The period from administration of the T cells in the dose described above to the onset of cranial nerve palsy is in the range of approximately 17 to approximately 60 days, and optionally, the median period from administration of the T cells in the dose described above to the onset of cranial nerve palsy is approximately 21 days; (3) The cranial nerve palsy occurs in the third, fifth, or fifth cranial nerve; (4) The duration of the cranial nerve palsy is in the range of approximately 15 days to approximately 262 days, and optionally, the median duration of the cranial nerve palsy is approximately 77 days; or (5) The treatment includes corticosteroids; (d) The CAR-T cell neurotoxicity is peripheral neuropathy, and optionally, peripheral neuropathy occurs in approximately 2.8% of the subjects; or (e) The CAR-T cell neurotoxicity is an adverse event that occurred during the experimental treatment for motor and neurocognitive function, and the adverse event that occurred during the experimental treatment for motor and neurocognitive function is of grade 1, and further, the adverse event of grade 1 that occurred during the experimental treatment for motor and neurocognitive function occurs in approximately 0.6% of the subjects, The method according to claim 62.

64. The method according to any one of claims 42 to 63, wherein the subject has one or more high-risk cytogenetic abnormalities selected from the group including Gain / amp(1q), del(17p), t(4;14), t(14;16), or any combination thereof.

65. The method according to claim 64, wherein the subject has at least two cytogenetic abnormalities, and optionally the subject has two, three, four, five, or more cytogenetic abnormalities.

66. The method according to claim 64 or 65, wherein the cytogenetic abnormality is a standard-risk cytogenetic abnormality.

67. The method according to any one of claims 1 to 66, wherein the administration of T cells in the aforementioned dose is effective in achieving a better-than-best partial response (VGPR) in the subject compared to the administration of DPd or PVd therapy.

68. The method according to claim 67, wherein approximately 81.3% of patients achieve a VGPR level of 2 or higher after administration of the aforementioned treatment.

69. The method according to claim 67 or 68, wherein approximately 45.5% of patients achieve a VGPR of 1 or higher after administration of DPd or PVd.

70. The method according to any one of claims 67 to 69, wherein the treatment is more effective in achieving a strict complete response (sCR) in the subject compared to the administration of DPd or PVd treatment.

71. The method according to claim 70, wherein the sCR after administration of the treatment is approximately 58.2%.

72. The method according to claim 70 or 71, wherein the sCR after administration of DPd or PVd is approximately 15.2%.

73. The method according to any one of claims 67 to 72, wherein administering the aforementioned dose of T cells reduces the risk of disease progression or death in the subject.

74. The method according to claim 73, wherein the risk of disease progression or death is reduced compared to the administration of daratumumab-pomalidomide-dexamethasone (DPd) or pomalidomide-bortezomib-dexamethasone (PVd) treatment.

75. The method according to claim 73, wherein the risk of disease progression or death is reduced compared to the administration of ide-cel therapy.

76. The method according to any one of claims 67 to 75, wherein the subject exhibits a disease progression or mortality risk reduced by approximately 60% to approximately 75%.

77. The method according to claim 76, wherein the subject exhibits a disease progression or mortality risk reduced by approximately 74%.

78. The method according to any one of claims 67 to 77, wherein the IMiD is lenalidomide.

79. The method according to any one of claims 67 to 78, wherein the subject has received three or fewer prior treatment lines.

80. The method according to any one of claims 67 to 79, wherein the subject has received two or fewer prior treatment lines.

81. The method according to any one of claims 67 to 80, wherein the subject has received only one prior treatment line.

82. The method according to any one of claims 67 to 81, which is effective in achieving an overall response rate (ORR) of approximately 75% to approximately 100%.

83. The method according to claim 82, wherein the ORR is approximately 84.6%.

84. The method according to any one of claims 67 to 83, wherein the treatment is effective in extending the median progression-free survival (PFS) of the subject compared to the administration of DPd or PVd treatment.

85. The method according to claim 84, wherein the PFS at 12 months after administration of the treatment is approximately 75.9%.

86. The method according to claim 84 or 85, wherein the PFS at 12 months after administration of DPd or PVd is approximately 48.6%.

87. The method further comprises administering the T cells in the dose described above and then treating the subject for CAR-T related adverse events, optionally comprising administering a treatment to the subject to mitigate the CAR-T related adverse events, optionally comprising the CAR-T related adverse events including cytokine release syndrome (CRS) and / or neurotoxicity, optionally, (a) The CAR-T-related adverse event is CRS, and optionally, the CRS occurs in approximately 60% to 90% or 76.1% of the subjects, and optionally, the maximum toxicity grade of the CRS is Grade 1, Grade 2 or Grade 3, and optionally, (1) The maximum toxicity grade of the CRS is Grade 1, and optionally, the maximum toxicity grade of Grade 1 occurs in approximately 52.8% of the subjects; (2) The maximum toxicity grade of the CRS is Grade 2, and optionally, the maximum toxicity grade of Grade 2 occurs in approximately 22.2% of the subjects; (3) The maximum toxicity grade of the CRS is Grade 3, and optionally, the maximum toxicity grade of Grade 3 occurs in approximately 1.1% of the subjects; (4) The period until the first onset of CRS is in the range of approximately 1 to approximately 23 days, and optionally, the median period until the first onset of CRS is approximately 8 days; (5) The duration of the CRS is in the range of approximately 1 to approximately 17 days, and optionally, the median duration of the CRS is approximately 3 days; or (6) The treatment includes tocilizumab, oxygen, corticosteroids, vasopressors, or any combination thereof; or (b) The CAR-T related adverse event is neurotoxic, and optionally, the neurotoxicity includes immunoeffector cell-associated neurotoxic syndrome or associated symptoms, motor / neurocognitive neurotoxicity, neurotoxic adverse events occurring under study treatment, non-immune effector cell-associated neurotoxic syndrome or associated symptoms, or any combination thereof, and optionally, the neurotoxicity is immunoeffector cell-associated neurotoxic syndrome or associated symptoms, and optionally, (1) The immunoeffector cell-associated neurotoxic syndrome or associated symptoms occur in approximately 4.5% of the subjects; (2) The maximum toxicity grade of the immunoeffector cell-associated neurotoxic syndrome or associated symptoms is Grade 1 or Grade 2, and optionally, the maximum toxicity grade of the immunoeffector cell-associated neurotoxic syndrome or associated symptoms is Grade 1, and optionally, the maximum toxicity grade of Grade 1 occurs in approximately 3.4% of the subjects, or optionally, the maximum toxicity grade of the immunoeffector cell-associated neurotoxic syndrome or associated symptoms is Grade 2, and optionally, the maximum toxicity grade of Grade 2 occurs in approximately 1.1% of the subjects; (3) The period until the onset of the immunoeffector cell-associated neurotoxic syndrome or associated symptoms is in the range of approximately 6 to approximately 15 days, and optionally, the median period until the onset of the immunoeffector cell-associated neurotoxic syndrome or associated symptoms is approximately 9.5 days; (4) The duration of the immunoeffector cell-associated neurotoxic syndrome or associated symptoms is in the range of approximately 1 to approximately 6 days, and optionally, the median duration of the immunoeffector cell-associated neurotoxic syndrome or associated symptoms is approximately 2 days; or (5) The treatment includes a corticosteroid and / or tocilizumab, The method according to any one of claims 67 to 86.

88. The neurotoxicity is CAR-T cell neurotoxicity, and optionally, the CAR-T cell neurotoxicity occurs in approximately 17.0% of the subjects, and optionally, the CAR-T cell neurotoxicity includes grade 3 / 4 neurotoxicity, grade 5 neurotoxicity, cranial nerve palsy, peripheral neuropathy, adverse events occurring during the study treatment relating to motor / neurocognitive function, or any combination thereof, and optionally, (a) The CAR-T cell neurotoxicity is a grade 3 / 4 neurotoxicity occurring in approximately 2.3% of the subjects; (b) The CAR-T cell neurotoxicity is Grade 5 neurotoxicity; (c) The CAR-T cell neurotoxicity is cranial nerve palsy, and in the subjects, the cranial nerve palsy occurs at a rate of approximately 9.1%, and in the subjects, (1) The cranial nerve palsy is of grade 2 or grade 3, and optionally, the cranial nerve palsy is of grade 2, occurring in approximately 8.0% of the subjects, and optionally, the cranial nerve palsy is of grade 3, occurring in approximately 1.1% of the subjects; (2) The period from administration of the T cells in the dose described above to the onset of cranial nerve palsy is in the range of approximately 17 to approximately 60 days, and optionally, the median period from administration of the T cells in the dose described above to the onset of cranial nerve palsy is approximately 21 days; (3) The cranial nerve palsy occurs in the third, fifth, or fifth cranial nerve; (4) The duration of the cranial nerve palsy is in the range of approximately 15 days to approximately 262 days, and optionally, the median duration of the cranial nerve palsy is approximately 77 days; or (5) The treatment includes corticosteroids; (d) The CAR-T cell neurotoxicity is peripheral neuropathy, and optionally, peripheral neuropathy occurs in approximately 2.8% of the subjects; or (e) The CAR-T cell neurotoxicity is an adverse event that occurred during the experimental treatment for motor and neurocognitive function, and the adverse event that occurred during the experimental treatment for motor and neurocognitive function is of grade 1, and further, the adverse event of grade 1 that occurred during the experimental treatment for motor and neurocognitive function occurs in approximately 0.6% of the subjects, The method according to claim 87.

89. The method according to any one of claims 67 to 88, wherein the subject has one or more high-risk cytogenetic abnormalities selected from the group including Gain / amp(1q), del(17p), t(4;14), t(14;16), or any combination thereof.

90. The method according to claim 89, wherein the subject has at least two cytogenetic abnormalities, and optionally the subject has two, three, four, five, or more cytogenetic abnormalities.

91. The method according to claim 89 or 90, wherein the cytogenetic abnormality is a standard-risk cytogenetic abnormality.

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