Therapy containing MTOR inhibitor nanoparticle composition

Abstract

In a particular embodiment, this application relates to a method and composition for treating cancer such as undifferentiated pleomorphic sarcoma or leiomyosarcoma using (a) a composition comprising nanoparticles containing an mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or a derivative thereof) and albumin, and optionally (b) an anti-PD-1 antibody.

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims priority to U.S. Provisional Patent Application No. 63 / 463,052, filed on 30 April 2023, the entirety of which is incorporated herein by reference for all purposes.

[0002] This application relates, in a particular embodiment, to a method and composition for treating undifferentiated pleomorphic sarcoma or leiomyosarcoma using a composition comprising an mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or a derivative thereof) and albumin-containing nanoparticles. [Background technology]

[0003] Progressive solid tumors are almost always fatal, and innovative therapies are urgently needed. Mammalian targets of rapamycin (mTOR) are conserved serine / threonine kinases that incorporate intracellular and extracellular signals and function as a central hub of signaling in cells to regulate cell proliferation and homeostasis. Activation of the mTOR pathway is associated with cell proliferation and survival, while inhibition of mTOR signaling is associated with inflammation and cell death. Dysregulation of the mTOR signaling pathway is involved in the increasing number of human diseases, including cancers such as progressive solid tumors and autoimmune diseases. Therefore, mTOR inhibitors have found broad applications in the treatment of diverse pathological conditions such as solid tumors, hematological malignancies, organ transplantation, restenosis, and rheumatoid arthritis.

[0004] Sirolimus (INN / USAN), also known as rapamycin, is an immunosuppressant used to prevent rejection in organ transplants, and is particularly useful in kidney transplants. Sirolimus-eluting stents are approved in the United States for the treatment of coronary artery restenosis. Furthermore, sirolimus has been demonstrated as an effective inhibitor of tumor growth in various cell lines and animal models. Other limus-based drugs, such as sirolimus analogs, have been designed to improve upon the pharmacokinetic and pharmacodynamic properties of sirolimus. For example, temsirolimus is approved in the United States and Europe for the treatment of renal cell carcinoma. Everolimus is approved in the United States for the treatment of advanced breast cancer, pancreatic neuroendocrine tumors, advanced renal cell carcinoma, and subependymal giant cell astrocytoma (SEGA) associated with tuberous sclerosis. Sirolimus acts by binding to the cytoplasmic protein FK-binding protein 12 (FKBP12), and the sirolimus-FKBP12 complex inhibits the mTOR pathway by directly binding to mTOR complex 1 (mTORC1).

[0005] Albumin-based nanoparticle compositions have been developed as drug delivery systems for delivering substantially water-insoluble drugs. See, for example, U.S. Patents 5,916,596, 6,506,405, 6,749,868, 6,537,579, 7,820,788, and 7,923,536. Abraxane®, an albumin-stabilized nanoparticle formulation of paclitaxel, was approved in the United States in 2005 and subsequently approved in various other countries for the treatment of metastatic breast cancer. It was recently approved in the United States for the treatment of non-small cell lung cancer and has demonstrated therapeutic efficacy in various clinical trials for the treatment of difficult-to-treat cancers such as bladder cancer and melanoma. Albumin derived from human blood is used in the manufacture of Abraxane® and various other albumin-based nanoparticle compositions. Albumin-based nanoparticle compositions containing sirolimus, for example, nab-sirolimus or Fyarrao®, are known, for example, U.S. Patent No. 8,911,786 and U.S. Patent No. 11,497,737.

[0006] However, in this field, there is still a need for advanced treatments for certain types of cancer. [Overview of the project]

[0007] In a particular embodiment, the present invention provides a method for treating undifferentiated pleomorphic sarcoma in an individual requiring treatment, the method comprising administering to the individual (a) an effective amount of a composition comprising nanoparticles containing an mTOR inhibitor and albumin, and optionally (b) an effective amount of a second therapeutic agent (e.g., an anti-PD-1 antibody).

[0008] In some embodiments, undifferentiated pleomorphic sarcoma has a loss of phosphatase tensin homolog (PTEN). In some embodiments, the PTEN loss is a loss-of-function mutation or epigenetic silencing. In some embodiments, individuals are selected for treatment based on having PTEN loss. In some embodiments, the method further includes selecting individuals based on having PTEN loss. In some embodiments, undifferentiated pleomorphic sarcoma has a tuberous sclerosis complex 2 (TSC2) mutation. In some embodiments, the TSC2 mutation is a missense mutation, a nonsense mutation, a deletion, a splicing site mutation, an insertion, a substation, a rearrangement, or a frameshift, or a combination thereof. In some embodiments, individuals are selected for treatment based on the TSC2 mutation. In some embodiments, the method further includes selecting individuals based on having the TSC2 mutation.

[0009] In another embodiment, the present invention provides a method for treating leiomyosarcoma in an individual requiring treatment, the method comprising administering to the individual (a) an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and albumin, and optionally (b) an effective amount of a second therapeutic agent (e.g., an anti-PD-1 antibody).

[0010] In some embodiments, the leiomyosarcoma is estrogen receptor-positive leiomyosarcoma. In some embodiments, individuals are selected for treatment based on having an estrogen receptor-positive leiomyosarcoma. In some embodiments, the method further includes selecting individuals based on having an estrogen receptor-positive leiomyosarcoma.

[0011] In some embodiments, undifferentiated pleomorphic sarcoma or leiomyosarcoma is locally progressive, progressive, malignant, progressively malignant, or metastatic. In some embodiments, undifferentiated pleomorphic sarcoma or leiomyosarcoma is relapsed or refractory to prior treatment. In some embodiments, prior treatment comprises a composition comprising an mTOR inhibitor and albumin-containing nanoparticles.

[0012] In some embodiments, the amount of mTOR inhibitor in the mTOR inhibitor nanoparticle composition is approximately 10 mg / m 2 ~about 150mg / m 2 In some embodiments, the amount of mTOR inhibitor in the mTOR inhibitor nanoparticle composition is approximately 100 mg / m 2 In some embodiments, the amount of mTOR inhibitor in the mTOR inhibitor nanoparticle composition is approximately 75 mg / m 2 In some embodiments, the amount of mTOR inhibitor in the mTOR inhibitor nanoparticle composition is approximately 56 mg / m 2 In some embodiments, the amount of mTOR inhibitor in the mTOR inhibitor nanoparticle composition is approximately 45 mg / m 2 In some embodiments, the amount of mTOR inhibitor in the mTOR inhibitor nanoparticle composition is approximately 30 mg / m 2 That is the case.

[0013] In some embodiments, the mTOR inhibitor nanoparticle composition is administered weekly. In some embodiments, the mTOR inhibitor nanoparticle composition is administered for two weeks every three weeks. In some embodiments, the mTOR inhibitor nanoparticle composition is administered on days 8 and 15 of a 21-day cycle.

[0014] In some embodiments, the mTOR inhibitor nanoparticle composition and the anti-PD-1 antibody are administered to the individual in parallel. In some embodiments, the mTOR inhibitor nanoparticle composition and the anti-PD-1 antibody are administered to the individual sequentially. In some embodiments, the mTOR inhibitor nanoparticle composition and the anti-PD-1 antibody are administered to the individual simultaneously.

[0015] In some embodiments, the second therapeutic agent (e.g., an anti-PD-1 antibody) is administered in an amount of approximately 1 mg / kg to approximately 10 mg / kg.

[0016] In some embodiments, the second therapeutic agent (e.g., an anti-PD-1 antibody) is administered every three weeks. In some embodiments, the second therapeutic agent (e.g., an anti-PD-1 antibody) is administered on day 1 of a 21-day cycle.

[0017] In some embodiments, a second therapeutic agent (e.g., an anti-PD-1 antibody) is administered for at least one cycle prior to the administration of the mTOR inhibitor nanoparticle composition.

[0018] In some embodiments, the mTOR inhibitor is a limus-based drug. In some embodiments, the limus-based drug is sirolimus.

[0019] In some embodiments, the average diameter of nanoparticles in the composition is about 150 nm or less. In some embodiments, the average diameter of nanoparticles in the composition is about 120 nm or less.

[0020] In some embodiments, the weight ratio of albumin to the mTOR inhibitor in the nanoparticle composition is approximately 9:1 or less.

[0021] In some embodiments, the nanoparticles contain an mTOR inhibitor associated with albumin. In some embodiments, the nanoparticles contain an mTOR inhibitor coated with albumin.

[0022] In some embodiments, the mTOR inhibitor nanoparticle composition is administered intravenously, intra-arterially, intraperitoneally, intravesically, subcutaneously, intrathecally, intrapulmonaryly, intramuscularly, intratracheally, intraocularly, transdermally, orally, or by inhalation. In some embodiments, the mTOR inhibitor nanoparticle composition is administered intravenously.

[0023] In some embodiments, the anti-PD-1 antibody is selected from the group consisting of nivolumab, pembrolizumab, semiprimab, atezolizumab, dostallimab, durvalumab, and avelumab. In some embodiments, the anti-PD-1 antibody is nivolumab.

[0024] In some embodiments, the individual is a human.

[0025] All publications, patents, patent applications, and disclosures of published patent applications referenced herein are incorporated herein by reference in their entirety. [Modes for carrying out the invention]

[0026] In certain embodiments, this application provides a treatment for cancers such as undifferentiated pleomorphic sarcoma (including undifferentiated pleomorphic sarcoma with PTEN loss and / or TSC2 mutation) or leiomyosarcoma (including estrogen receptor-positive leiomyosarcoma), comprising (a) a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or a derivative thereof) and albumin (e.g., nab-sirolimus), and optionally, (b) a second therapeutic agent (e.g., an anti-PD-1 antibody (e.g., nivolumab)). In some embodiments, the method is for the treatment of undifferentiated pleomorphic sarcoma with PTEN loss and / or TSC2 mutation in an individual requiring treatment. In some embodiments, the method is for the treatment of estrogen receptor-positive leiomyosarcoma in an individual requiring treatment.

[0027] A particular subject of this application is, at least in part, based on the finding that individuals with undifferentiated pleomorphic sarcoma (including undifferentiated pleomorphic sarcoma with PTEN loss and / or TSC2 mutation) or leiomyosarcoma (including estrogen receptor-positive leiomyosarcoma) who received combination therapy unexpectedly showed an improved response to treatment with nab-sirolimus and the anti-PD-1 antibody nivolumab compared to individuals with other cancer subtypes.

[0028] Accordingly, in a particular embodiment, a method is provided for treating an undifferentiated pleomorphic sarcoma having PTEN loss and / or TSC2 mutation (e.g., PTEN loss and TSC2 mutation) in an individual requiring treatment, the method comprising administering to the individual (a) an effective amount of a composition comprising nanoparticles containing an mTOR inhibitor and albumin, e.g., nab-sirolimus, and optionally, (b) a second therapeutic agent (e.g., an effective amount of an anti-PD-1 antibody, e.g., nivolumab).

[0029] In another embodiment, this specification provides a method for treating estrogen receptor-positive leiomyosarcoma in an individual in need of treatment, the method comprising administering to the individual (a) an effective amount of a composition comprising nanoparticles containing an mTOR inhibitor and albumin, e.g., nab-sirolimus, and optionally (b) an effective amount of a second therapeutic agent (e.g., an anti-PD-1 antibody, e.g., nivolumab).

[0030] I. Definition As used herein, “nab” stands for nanoparticle albumin binding, and “nab-sirolimus” is an albumin-stabilized nanoparticle formulation of sirolimus. Nab-sirolimus is also known as nab-rapamycin, as described above. See, for example, U.S. Patent Nos. 8,911,786 and 11,497,737, each of which is incorporated herein by reference as a whole.

[0031] As used herein, “treatment” or “treating” refers to an approach to obtain beneficial or desired outcomes, including clinical outcomes. For the purposes of the present invention, beneficial or desired clinical outcomes include, but are not limited to, one or more of the following: relief of one or more symptoms caused by the disease; reduction of the scope of the disease; stabilization of the disease (e.g., prevention or delay of disease exacerbation); prevention or delay of disease spread (e.g., metastasis); prevention or delay of disease recurrence; reduction of disease recurrence rate; delay or delay of disease progression; improvement of disease status; provision of disease remission (partial or total); reduction of the dose of one or more other drugs required to treat the disease; delay of disease progression; improvement of quality of life; and / or extension of survival. In some embodiments, treatment reduces the severity of one or more symptoms associated with cancer by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 100% compared to the corresponding symptoms in the same subject before treatment, or compared to the corresponding symptoms in other untreated subjects. The reduction of pathological outcomes of cancer is also included in "treatment." The method of the present invention aims to achieve any one or more of these aspects of treatment.

[0032] The terms "recurrence," "relapse," or "relapsed" refer to the recurrence of cancer or disease after the disease has been clinically evaluated as cleared. A diagnosis of distant metastasis or local recurrence may be considered a recurrence.

[0033] The terms "refractory" or "resistant" refer to cancer or disease that has not responded to treatment.

[0034] As used herein, “delaying” the onset of cancer means postponing, interfering with, slowing, delaying, stabilizing, and / or postponing the onset of the disease. This delay may be of varying duration depending on the medical history and / or the individual being treated. As will be apparent to those skilled in the art, sufficient or significant delay may substantially encompass prevention, in the sense that the individual does not develop the disease. A method of “delaying” the onset of cancer is a method that reduces the likelihood of developing the disease in a particular time frame and / or reduces the severity of the disease in a particular time frame compared to not using the method. Such comparisons are typically based on clinical studies using a statistically significant number of subjects. Cancer onset can be detected using standard methods, including, but not limited to, computed tomography (CAT scan), magnetic resonance imaging (MRI), ultrasound, coagulation tests, angiography, biopsy, urinalysis, and cystoscopy. Onset may also refer to the progression of cancer, including development, recurrence, and onset, and may initially be undetectable.

[0035] As used herein, the term “effective dose” refers to an amount of a compound or composition sufficient to treat a particular disorder, condition, or disease, for example, to improve, alleviate, reduce, and / or delay one or more of its symptoms. With respect to cancer, an effective dose includes an amount sufficient to shrink a tumor and / or reduce its growth rate (such as inhibiting tumor growth), or to prevent or delay other unwanted cell proliferation in the cancer. In some embodiments, an effective dose is sufficient to delay the onset of cancer. In some embodiments, an effective dose is sufficient to prevent or delay recurrence. In some embodiments, an effective dose is sufficient to reduce the recurrence rate in an individual. An effective dose may be administered in one or more doses. An effective amount of drug or composition may (i) reduce the number of cancer cells, (ii) reduce tumor size, (iii) inhibit, suppress, to some extent slow down, preferably stop, the invasion of cancer cells into peripheral organs, (iv) inhibit tumor metastasis (i.e., to some extent slow down, preferably stop), (v) inhibit tumor growth, (vi) prevent or delay tumor development and / or recurrence, (vii) reduce the recurrence rate of tumors, and / or (viii) alleviate to some extent one or more of the symptoms associated with cancer.

[0036] As is understood in the art, an effective dose may be one or more doses, i.e., a single dose or multiple doses may be required to achieve the desired therapeutic endpoint. An effective dose may be considered in the context of administering one or more therapeutic agents, and a nanoparticle composition (e.g., a composition comprising sirolimus and albumin) may be considered to be given in an effective dose if, in combination with one or more other agents, a desirable or beneficial outcome can be achieved, or if such an outcome is achieved. The components in the combination therapy of the present invention (e.g., the first and second therapies) may be administered sequentially, simultaneously, or in parallel using the same or different routes of administration for each component. Thus, an effective dose of combination therapy includes the amount of the first therapy and the amount of the second therapy that, when administered sequentially, simultaneously, or in parallel, yield the desired outcome.

[0037] "In conjunction with" or "in combination with" means administering one treatment in addition to another, for example, administering the nanoparticle composition described herein in addition to administering other drugs to the same individual under the same treatment plan. Therefore, "in conjunction with" or "in combination with" means administering one treatment before, during, or after the delivery of other treatments to the individual.

[0038] As used herein, the term "simultaneous administration" means that the first and second therapeutic agents in a combination therapy are administered with an interval of about 15 minutes or less, for example, about 10 minutes or less, 5 minutes or less, or 1 minute or less. When the first and second therapies are administered simultaneously, the first and second therapies may be contained in the same composition (e.g., a composition containing both the first and second therapeutic agents) or in separate compositions (e.g., one composition containing the first therapy and another containing the second therapy).

[0039] As used herein, the term “sequential administration” means that the first and second therapies in combination therapy are administered at intervals of more than about 15 minutes, for example, more than about 20 minutes, more than 30 minutes, more than 40 minutes, more than 50 minutes, more than 60 minutes, or longer. Either the first or second therapy may be administered first. The first and second therapies are contained in separate compositions, which may be contained in the same or different packaging or kits.

[0040] As used herein, “concurrent administration” means that the administration of the first therapeutic agent and the administration of the second therapeutic agent in combination therapy overlap with each other.

[0041] The term "antibody" includes full-length antibodies and their antigen-binding fragments. A full-length antibody comprises two heavy chains and two light chains. Variable regions of the light and heavy chains are involved in antigen binding. The variable regions of both chains generally comprise three highly variable loops called complementarity-determining regions (CDRs): the light chain (LC)CDRs containing LC-CDR1, LC-CDR2, and LC-CDR3, and the heavy chain (HC)CDRs containing HC-CDR1, HC-CDR2, and HC-CDR3. The CDR boundaries for antibodies and antigen-binding fragments disclosed herein may be defined or identified by the rules of Kabat, Chothia, or Al-Lazikani (Al-Lazikani 1997; Chothia 1985; Chothia 1987; Chothia 1989; Kabat 1987; Kabat 1991). Three CDRs of the heavy or light chain are inserted between adjacent stretches known as framework regions (FRs), which are more conserved than the CDRs and form a scaffold supporting the hypervariable loop. The constant regions of the heavy and light chains do not participate in antigen binding but exhibit various effector functions. Antibodies are assigned to classes based on the amino acid sequence of the constant region of their heavy chains. The five major classes or isotypes of antibodies are IgA, IgD, IgE, IgG, and IgM, characterized by the presence of α, δ, ε, γ, and μ heavy chains, respectively. Some major antibody classes are classified into subclasses such as IgG1 (γ1 heavy chain), IgG2 (γ2 heavy chain), IgG3 (γ3 heavy chain), IgG4 (γ4 heavy chain), IgGA1 (α1 heavy chain), or IgGA2 (α2 heavy chain).

[0042] As used herein, the term “antigen-binding fragment” refers to an antibody fragment that includes, for example, a diabody, Fab, Fab', F(ab')2, Fv fragment, disulfide-stabilized Fv fragment (dsFv), (dsFv)2, bispecific dsFv (dsFv-dsFv'), disulfide-stabilized diabody (dsdiabody), single-chain antibody molecule (scFv), scFv dimer (bivalent diabody), a multispecific antibody formed from a portion of an antibody containing one or more CDRs, a camelized single-domain antibody, a nanobody, a domain antibody, a bivalent domain antibody, or any other antibody fragment that binds to an antigen but does not contain a complete antibody structure. An antigen-binding fragment can bind to the same antigen to which a parent antibody or parent antibody fragment (e.g., parent scFv) binds. In some embodiments, an antigen-binding fragment may include one or more CDRs from a particular human antibody transplanted into a framework region from one or more different human antibodies.

[0043] As used herein, the terms “specifically binding” or “specific to” refer to a measurable and reproducible interaction, such as binding between a target and an antibody or antibody moiety, which determines the presence of the target in the presence of a heterogeneous population of molecules, including biological molecules. For example, an antibody or antibody moiety that specifically binds to a target (which may be an epitope) is an antibody or antibody moiety that binds to this target more readily and / or for a longer duration with greater affinity, avidity, than to other targets. In some embodiments, an antibody or antibody moiety that specifically binds to an antigen reacts with one or more antigenic determinants of an antigen (e.g., PD-1 or a portion thereof) with a binding affinity that is at least about 10 times its binding affinity to other targets.

[0044] As used herein, “pharmaceutically acceptable” or “pharmacologically compatible” means a material that is not biologically or otherwise undesirable, for example, a material that can be incorporated into a pharmaceutical composition administered to a patient without causing any significant undesirable biological effects or any adverse interactions with any of the other components of the composition in which it is contained. Pharmacologically acceptable carriers or excipients preferably meet the required criteria for toxicity and manufacturing testing and / or are listed in the non-active ingredient guide prepared by the U.S. Food and Drug Administration.

[0045] As used herein, the term “individual” refers to mammals, including but not limited to humans, cattle, horses, cats, dogs, rodents, rats, mice, or primates. In some embodiments, the individual is a human individual.

[0046] The terms “comprising,” “having,” “containing,” and “including,” as well as other similar forms and their grammatical equivalents, are intended to be semantically equivalent and open-ended, in that the one or more items following any one of these words do not mean an exhaustive list of such one or more items, or that they are limited to only the listed one or more items. For example, an article “containing” components A, B, and C may consist of (i.e., contain only) components A, B, and C, or it may include not only components A, B, and C but also one or more other components. As such, “~containing” and similar forms and their grammatical equivalents are intended and understood to include disclosures of embodiments of “~essentially consisting of” or “~ comprising.”

[0047] Where a range of values ​​is provided, unless the context explicitly indicates otherwise, the values ​​between the upper and lower limits of that range, and the intermediate values ​​up to one-tenth of the lower limit in any other stated or intermediate values ​​within that stated range, are included within the scope of this disclosure and are subject to any restrictions specifically excluded within the stated scope. If the stated scope includes one or both limits, the scope excluding one or both of those included limits is also included in this disclosure.

[0048] In this specification, any reference to a value or parameter using the term "about" includes (and describes) variations relating to the value or parameter itself. For example, any reference to "about X" includes a description of "X".

[0049] As used herein, including in the attached claims, the singular “a,” “or,” and “the” refer to multiple subjects unless otherwise explicitly stated in the context.

[0050] Those skilled in the art will recognize that several embodiments are possible within the scope and spirit of this disclosure. The following description is illustrative of this disclosure and should not be construed as limiting the scope of the inventions described herein.

[0051] II. Treatment method This specification provides a method for treating cancers, including undifferentiated pleomorphic sarcomas (including undifferentiated pleomorphic sarcomas with PTEN loss and / or TSC2 mutations) or leiomyosarcomas (including estrogen receptor-positive leiomyosarcomas), in individuals in need of treatment, the treatment comprising administering to the individual (a) a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or a derivative thereof) and albumin, and optionally (b) a second therapeutic agent (e.g., an anti-PD-1 antibody).

[0052] In some embodiments, a method is provided for treating undifferentiated pleomorphic sarcoma having PTEN loss and / or TSC2 mutations in an individual requiring treatment, the treatment comprising administering to the individual (a) a composition comprising nanoparticles containing an mTOR inhibitor (limus-based drug, e.g., sirolimus or a derivative thereof) and albumin (e.g., nab-sirolimus), and optionally, (b) a second therapeutic agent (e.g., an anti-PD-1 antibody (e.g., nivolumab)). In some embodiments, PTEN loss is a loss-of-function mutation or epigenetic silencing. In some embodiments, the individual is selected for treatment based on having PTEN loss. In some embodiments, the TSC2 mutation is a missense mutation, a nonsense mutation, a deletion, a splicing site mutation, an insertion, a substation, a rearrangement, or a frameshift, or a combination thereof. In some embodiments, the individual is selected for treatment based on the TSC2 mutation. In some embodiments, the method further comprises selecting the individual based on having PTEN loss and / or a TSC2 mutation.

[0053] In some embodiments, a method is provided for treating estrogen receptor-positive leiomyosarcoma in an individual requiring treatment, the treatment comprising administering to the individual (a) a composition comprising nanoparticles containing an mTOR inhibitor (limus-based drug, e.g., sirolimus or a derivative thereof) and albumin (e.g., nab-sirolimus), and optionally (b) a second therapeutic agent (e.g., an anti-PD-1 antibody (e.g., nivolumab)).

[0054] In some embodiments, the cancer, such as undifferentiated pleomorphic sarcoma or leiomyosarcoma, is locally advanced, progressive, malignant, progressively malignant, or metastatic. In some embodiments, the cancer, such as undifferentiated pleomorphic sarcoma or leiomyosarcoma, is recurrent, refractory, or resistant to prior treatment. In some embodiments, the prior treatment comprises a composition comprising nanoparticles containing an mTOR inhibitor and albumin, e.g., nab-sirolimus. In some embodiments, the prior treatment comprises a non-nanoparticle formulation of an mTOR inhibitor such as sirolimus.

[0055] The methods provided herein are applicable to all stages of cancer, including stages I, II, III, and IV, as defined by the American Joint Committee on Cancer (AJCC) staging groups, such as undifferentiated pleomorphic sarcoma or leiomyosarcoma. In some embodiments, the cancer, such as undifferentiated pleomorphic sarcoma or leiomyosarcoma, is early-stage cancer, non-metastatic cancer, primary cancer, advanced cancer, locally advanced cancer, metastatic cancer, cancer in remission, adjuvant-set cancer, or neoadjuvant-set cancer. In some embodiments, the solid tumor is locally resectable, locally unresectable, or unresectable. In some embodiments, the cancer, such as undifferentiated pleomorphic sarcoma or leiomyosarcoma, is locally resectable or borderline resectable.

[0056] A. Medication and administration methods The dose of an mTOR inhibitor nanoparticle composition (e.g., a sirolimus / albumin nanoparticle composition) administered to an individual (e.g., a human) may vary depending on the specific composition, the method of administration, and the specific stage of the tumor being treated. The amount should be sufficient to produce the desired response, such as a therapeutic or prophylactic response to the tumor. In some embodiments, the amount of the mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or its derivatives) in the composition is below the level that induces a toxic effect (e.g., an effect exceeding clinically acceptable toxicity levels) or at a level that allows for the control or tolerance of potential side effects when the mTOR inhibitor nanoparticle composition is administered to an individual.

[0057] In some embodiments, an mTOR inhibitor nanoparticle composition (e.g., a sirolimus / albumin nanoparticle composition) is administered to an individual simultaneously with a second therapeutic agent, such as an anti-PD-1 antibody. For example, the mTOR inhibitor nanoparticle composition and the anti-PD-1 antibody are administered at time intervals of approximately 15 minutes or less, for example, approximately 10, 5, or 1 minute or less. In one example, where the compounds are in solution, simultaneous administration can be achieved by administering a solution containing the combination of compounds. In another example, separate solutions can be administered simultaneously, one containing an mTOR inhibitor nanoparticle composition (e.g., a sirolimus / albumin nanoparticle composition) and the other containing an anti-PD-1 antibody. In one example, simultaneous administration can be achieved by administering a composition containing the combination of compounds. In yet another example, simultaneous administration can be achieved by administering two separate compositions, one containing an mTOR inhibitor nanoparticle composition (e.g., a sirolimus / albumin nanoparticle composition) and the other containing an anti-PD-1 antibody. In some embodiments, the co-administration of an mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or its derivatives) in a nanoparticle composition with an anti-PD-1 antibody can be combined with an adjunct dose of the mTOR inhibitor and / or anti-PD-1 antibody.

[0058] In other embodiments, the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition) and a second therapeutic agent, such as an anti-PD-1 antibody, are not administered simultaneously. In some embodiments, the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition) is administered before the anti-PD-1 antibody. In other embodiments, the anti-PD-1 antibody is administered before the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition). The time difference in non-simultaneous administration can be 1 minute, 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 2 hours, 3 hours, 6 hours, 9 hours, 12 hours, 24 hours, 36 hours, or more than 48 hours. In other embodiments, the first administered compound is provided with enough time to act on the patient before the second administered compound is administered. In some embodiments, the time difference does not extend beyond the time it takes for the first administered compound to complete its effect in the patient, or beyond the time it takes for the first administered compound to be completely or substantially eliminated or inactivated in the patient.

[0059] In some embodiments, the administration of an mTOR inhibitor nanoparticle composition (e.g., a sirolimus / albumin nanoparticle composition) and a second therapeutic agent such as an anti-PD-1 antibody is simultaneous, i.e., the administration periods of the mTOR inhibitor nanoparticle composition and the anti-PD-1 antibody overlap. In some embodiments, the mTOR inhibitor nanoparticle composition (e.g., a sirolimus / albumin nanoparticle composition) is administered for at least one cycle (e.g., at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 cycles) prior to the administration of the anti-PD-1 antibody. In some embodiments, the anti-PD-1 antibody is administered for at least 1, 2, 3, or 4 weeks. In some embodiments, the administration of the mTOR inhibitor nanoparticle composition (e.g., a sirolimus / albumin nanoparticle composition) and the anti-PD-1 antibody is initiated approximately simultaneously (e.g., within one of 1, 2, 3, 4, 5, 6, or 7 days). In some embodiments, the administration of an mTOR inhibitor nanoparticle composition (e.g., a sirolimus / albumin nanoparticle composition) and an anti-PD-1 antibody is terminated approximately simultaneously (e.g., within one of 1, 2, 3, 4, 5, 6, or 7 days). In some embodiments, the administration of the anti-PD-1 antibody continues after the termination of the administration of the mTOR inhibitor nanoparticle composition (e.g., a sirolimus / albumin nanoparticle composition) (e.g., for any of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months). In some embodiments, the administration of the anti-PD-1 antibody is initiated after the initiation of the administration of the mTOR inhibitor nanoparticle composition (e.g., a sirolimus / albumin nanoparticle composition) (e.g., after any of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months). In some embodiments, the administration of an mTOR inhibitor nanoparticle composition (e.g., a sirolimus / albumin nanoparticle composition) and an anti-PD-1 antibody is initiated and terminated approximately simultaneously.In some embodiments, the administration of an mTOR inhibitor nanoparticle composition (e.g., a sirolimus / albumin nanoparticle composition) and an anti-PD-1 antibody is initiated almost simultaneously, and the administration of the anti-PD-1 antibody continues after the discontinuation of the administration of the mTOR inhibitor nanoparticle composition (e.g., for about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months). In some embodiments, the administration of an mTOR inhibitor nanoparticle composition (e.g., a sirolimus / albumin nanoparticle composition) and an anti-PD-1 antibody is discontinued almost simultaneously, and the administration of the anti-PD-1 antibody is initiated after the start of the administration of the mTOR inhibitor nanoparticle composition (e.g., after about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months).

[0060] In some embodiments, a second therapeutic agent, such as an anti-PD-1 antibody, is administered on day 1 of a 21-day cycle, and the mTOR inhibitor nanoparticle composition is administered on days 8 and 15 of the 21-day cycle. In some embodiments, the first cycle comprises, for example, only the administration of the anti-PD-1 antibody on day 1. In some embodiments, the administration of the composition containing the mTOR inhibitor and albumin, along with the anti-PD-1 antibody, continues for at least one of the 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 cycles.

[0061] In some embodiments, the administration of an mTOR inhibitor nanoparticle composition (e.g., a sirolimus / albumin nanoparticle composition) and a second therapeutic agent, such as an anti-PD-1 antibody, is asynchronous. For example, in some embodiments, the administration of the mTOR inhibitor nanoparticle composition (e.g., a sirolimus / albumin nanoparticle composition) is completed before the administration of the anti-PD-1 antibody. In some embodiments, the administration of the anti-PD-1 antibody is completed before the administration of the mTOR inhibitor nanoparticle composition (e.g., a sirolimus / albumin nanoparticle composition). The period between these two asynchronous administrations can range from about 2 to 8 weeks, for example, about 4 weeks.

[0062] The dosing frequency of an mTOR inhibitor nanoparticle composition (e.g., a sirolimus / albumin nanoparticle composition) and a second therapeutic agent, such as an anti-PD-1 antibody, may be adjusted throughout the course of treatment at the discretion of the administering physician. When administered separately, the mTOR inhibitor nanoparticle composition (e.g., a sirolimus / albumin nanoparticle composition) and the second therapeutic agent, such as an anti-PD-1 antibody, can be administered at different dosing frequencies or intervals. For example, the mTOR inhibitor nanoparticle composition (e.g., a sirolimus / albumin nanoparticle composition) may be administered weekly, while the anti-PD-1 antibody may be administered at a more or less frequent frequency. In some embodiments, sustained-release formulations of nanoparticles and / or anti-PD-1 antibodies may be used. Various formulations and devices for achieving sustained release are known in the art. Combinations of dosing configurations described herein may also be used.

[0063] The mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition) and a second therapeutic agent, such as an anti-PD-1 antibody, can be administered using the same or different routes of administration. In some embodiments (both concurrent and sequential administration), the mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or its derivatives) and the anti-PD-1 antibody in the mTOR inhibitor nanoparticle composition are administered in a predetermined ratio.

[0064] The dose required for a second therapeutic agent, such as an mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or its derivatives) and / or an anti-PD-1 antibody, in an mTOR inhibitor nanoparticle composition may be the same as, or lower than, the dose typically required when each agent is administered alone (but not necessarily). Therefore, in some embodiments, an amount less than or equal to a therapeutic dose of the mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or its derivatives) and / or anti-PD-1 antibody in the mTOR inhibitor nanoparticle composition is administered. “Amount less than or equal to a therapeutic dose” or “level less than or equal to a therapeutic dose” refers to an amount less than a therapeutic dose, i.e., less than the amount typically used when the mTOR inhibitor nanoparticle composition (e.g., a sirolimus / albumin nanoparticle composition) and / or anti-PD-1 antibody is administered alone. The reduction may be reflected in the amount administered in a given dose and / or the amount administered over a given period of time (reduced frequency).

[0065] In some embodiments, a sufficient amount of a second therapeutic agent (e.g., an anti-PD-1 antibody) is administered to allow for a reduction in the usual dose of the mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or a derivative thereof) in the mTOR inhibitor nanoparticle composition, which is necessary to have an effect of at least about 5%, 10%, 20%, 30%, 50%, 60%, 70%, 80%, 90%, or more on the same level of treatment.

[0066] In some embodiments, the doses of both the mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or its derivatives) and the anti-PD-1 antibody in the mTOR inhibitor nanoparticle composition are reduced compared to their respective corresponding normal doses when administered alone. In some embodiments, both the mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or its derivatives) and the anti-PD-1 antibody in the mTOR inhibitor nanoparticle composition are administered at sub-thermal levels, i.e., reduced levels. In some embodiments, the doses of the mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or its derivatives) and / or the anti-PD-1 antibody in the mTOR inhibitor nanoparticle composition are substantially less than the established maximum toxic dose (MTD). For example, the doses of the mTOR inhibitor nanoparticle composition (e.g., a sirolimus / albumin nanoparticle composition) and / or the anti-PD-1 antibody are less than about 50%, 40%, 30%, 20%, or 10% of the MTD.

[0067] The combinations of dosage forms described herein may be used. The combination therapies described herein may be performed alone or in combination with other therapies such as surgery, radiation, gene therapy, immunotherapy, bone marrow transplantation, stem cell transplantation, hormone therapy, targeted therapy, cryotherapy, ultrasound therapy, photodynamic therapy, and / or chemotherapy. In addition, individuals at higher risk of developing solid tumors may receive treatments that inhibit and / or delay the onset of the disease.

[0068] As those skilled in the art will understand, in some embodiments, the appropriate dose of the second agent is approximately one already used in clinical therapy in which the anti-PD-1 antibody is administered alone or in combination with other chemotherapeutic agents. Dose variations occur depending on the condition being treated. As described above, in some embodiments, the second chemotherapeutic agent may be administered at reduced levels.

[0069] In some embodiments, the amounts of the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition) and the anti-PD-1 antibody are less than the levels that induce a toxic effect (i.e., an effect exceeding clinically acceptable toxicity levels), or are at levels that can control or tolerate potential side effects when the mTOR inhibitor nanoparticle composition and the anti-PD-1 antibody are administered to an individual.

[0070] In some embodiments, the amount of the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition) is close to the maximum tolerated dose (MTD) of the composition following the same dosing schedule when administered with the anti-PD-1 antibody. In some embodiments, the amount of the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition) is greater than any of about 80%, 90%, 95% or 98% of the MTD when administered with the anti-PD-1 antibody.

[0071] In some embodiments, the amount of the mTOR inhibitor (e.g., a rapamycin-based drug, e.g., sirolimus) in the mTOR inhibitor nanoparticle composition is about 25 mg / m 2 、30 mg / m 2 、45 mg / m 2 、50 mg / m 2 、56 mg / m 2 、60 mg / m 2 、75 mg / m 2 、80 mg / m 2 、90 mg / m 2 、100 mg / m 2 、120 mg / m 2 、160 mg / m 2 、175 mg / m 2 、180 mg / m 2 、200 mg / m 2 、210 mg / m 2 、220 mg / m 2 、250 mg / m 2 、260 mg / m 2 、300 mg / m 2 、350 mg / m 2 、400 mg / m 2 、500 mg / m 2 、540 mg / m2 750 mg / m² 2 , 1000 mg / m² 2 Or 1080 mg / m² 2 It is one of the mTOR inhibitors. In some embodiments, the mTOR inhibitor nanoparticle composition is approximately 350 mg / m 2 , 300 mg / m² 2 , 250 mg / m² 2 , 200 mg / m² 2 , 150 mg / m² 2 , 120 mg / m² 2 , 100 mg / m² 2 90 mg / m² 2 50 mg / m² 2 , or 30 mg / m² 2 It contains less than any of the following mTOR inhibitors (e.g., limus-based drugs, e.g., sirolimus). In some embodiments, the amount of mTOR inhibitor (e.g., limus-based drugs, e.g., sirolimus) per single dose is about 25 mg / m² 2 , 22 mg / m² 2 , 20 mg / m² 2 , 18 mg / m² 2 , 15 mg / m² 2 , 14 mg / m² 2 , 13 mg / m² 2 , 12 mg / m² 2 , 11 mg / m² 2 , 10 mg / m² 2 , 9 mg / m² 2 , 8 mg / m² 2 , 7 mg / m² 2 , 6 mg / m² 2 , 5 mg / m² 2 , 4 mg / m² 2 , 3 mg / m² 2 , 2 mg / m² 2 , or 1 mg / m² 2 It is less than any of the following. In some embodiments, the effective amount of the mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus) in the mTOR inhibitor nanoparticle composition is about 1 to about 5 mg / m 2 , about 5 to about 10 mg / m 2 , about 10 to about 25mg / m 2 , about 25~50mg / m 2 , about 50 to about 75mg / m 2, about 75~100mg / m 2 , about 100~125mg / m 2 , about 125~150mg / m 2 , about 150~175mg / m 2 , about 175~200mg / m 2 , about 200~225mg / m 2 , about 225~250mg / m 2 , about 250~300mg / m 2 , about 300~350mg / m 2 , or approximately 350-400 mg / m² 2 It falls within one of the following ranges. In some embodiments, the effective amount of the mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus) in the mTOR inhibitor nanoparticle composition is about 30 to about 300 mg / m 2 For example, approximately 100 to 150 mg / m² 2 , about 120mg / m2, about 130mg / m 2 , or approximately 140 mg / m² 2 In some embodiments, this amount of mTOR inhibitor nanoparticle composition is administered weekly. In some embodiments, this amount of mTOR inhibitor nanoparticle composition is administered weekly every two weeks out of three weeks. In some embodiments, this amount of mTOR inhibitor nanoparticle composition is administered on days 8 and 15 of a 21-day cycle, day 1 or 8 of a 21-day cycle, day 15 and 21 of a 21-day cycle, day 1 and 15 of a 21-day cycle, or day 1 and 21 of a 21-day cycle.

[0072] In some embodiments, the anti-PD-1 antibody is selected from the group consisting of nivolumab, pembrolizumab, semiprimab, atezolizumab, dostallimab, durvalumab, and avelumab. In some embodiments, the amount of anti-PD-1 antibody (e.g., nivolumab) is about 1 mg / kg to about 10 mg / kg. In some embodiments, the amount of anti-PD-1 antibody (e.g., nivolumab) is at least about 1 mg / kg, for example, at least about 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, or 10 mg / kg. In some embodiments, the dose of anti-PD-1 antibody (e.g., nivolumab) is 1 mg / kg or less, for example, 2 mg / kg or less, 3 mg / kg or less, 4 mg / kg or less, 5 mg / kg or less, 6 mg / kg or less, 7 mg / kg or less, 8 mg / kg or less, 9 mg / kg or less, or 10 mg / kg or less. In some embodiments, the dose of anti-PD-1 antibody (e.g., nivolumab) is approximately 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, or 10 mg / kg. In some embodiments, this dose of anti-PD-1 antibody is administered weekly, every two weeks, every three weeks, or monthly. In some embodiments, this dose of anti-PD-1 antibody is administered every three weeks. In some embodiments, this dose of anti-PD-1 antibody is administered every 21 days.

[0073] In some embodiments, the dosage frequency for administering the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition) may be daily, every two days, every three days, every four days, every five days, every six days, weekly without interruption, for three weeks out of four (e.g., days 1, 8, and 15 of a 28-day cycle), once every three weeks, once every two weeks, or for two weeks out of three weeks. In some embodiments, the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition) may be administered approximately once every two weeks, once every three weeks, once every four weeks, once every six weeks, or once every eight weeks. In some embodiments, the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition) may be administered at least approximately once, twice, three times, four times, five times, six times, or seven times per week (i.e., daily). In some embodiments, the interval between doses is less than one of the following: approximately 6 months, 3 months, 1 month, 20 days, 15 days, 14 days, 13 days, 12 days, 11 days, 10 days, 9 days, 8 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or 1 day. In some embodiments, the interval between doses is more than approximately 1 month, more than 2 months, more than 3 months, more than 4 months, more than 5 months, more than 6 months, more than 8 months, or more than 12 months. In some embodiments, there is no interruption in the medication schedule. In some embodiments, the interval between doses is approximately 1 week or less.

[0074] In some embodiments, the dosing frequency is once every two days for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 doses. In some embodiments, the dosing frequency is once every two days for 5 doses. In some embodiments, the mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or its derivatives) is administered over a period of at least 10 days, with intervals between each administration of approximately 2 days or less, and the dose of the mTOR inhibitor at each administration is approximately 0.25 mg / m². 2 ~about 250mg / m 2 , about 0.25mg / m 2 ~about 150mg / m 2 , about 0.25mg / m2 ~about 75 mg / m 2 、for example, about 0.25 mg / m 2 ~about 25 mg / m 2 、or about 25 mg / m 2 ~about 50 mg / m 2 is.

[0075] Administration of the mTOR inhibitor nanoparticle composition (for example, sirolimus / albumin nanoparticle composition) can be extended over a long period such as about 1 month to about 7 years. In some embodiments, the mTOR inhibitor nanoparticle composition (for example, sirolimus / albumin nanoparticle composition) is administered over any period of at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 48, 60, 72 or 84 months.

[0076] In some embodiments, the dosage of the mTOR inhibitor (for example, a rapamycin-based drug, for example, sirolimus or a derivative thereof) in the nanoparticle composition is 5 - 400 mg / m when given on a 3-week schedule 2 、or 5 - 250 mg / m when given on a weekly schedule 2 (for example, 80 - 150 mg / m 2 、for example, 100 - 120 mg / m 2 ) can be in the range. For example, the amount of the mTOR inhibitor (for example, a rapamycin-based drug, for example, sirolimus or a derivative thereof) is about 60 - about 300 mg / m on a 3-week schedule 2 (for example, about 260 mg / m 2 ).

[0077] In some embodiments, as an exemplary dosing schedule for administration of the mTOR inhibitor nanoparticle composition (for example, sirolimus / albumin nanoparticle composition), 100 mg / m weekly without interruption 2 、10 mg / m weekly for 3 out of 4 weeks (for example, on days 1, 8 and 15 of a 28-day cycle) 2 、45 mg / m weekly for 3 out of 4 weeks (for example, on days 1, 8 and 15 of a 28-day cycle) 275 mg / m² weekly for 3 weeks out of 4 weeks (for example, days 1, 8, and 15 of a 28-day cycle). 2 100 mg / m² weekly for 3 weeks out of 4 weeks. 2 125 mg / m² per week for 3 weeks out of 4 weeks. 2 125 mg / m² weekly for two weeks out of three. 2 130 mg / m² per week without interruption. 2 175 mg / m² once every two weeks. 2 260 mg / m² once every two weeks. 2 260 mg / m² once every three weeks. 2 180-300 mg / m² every three weeks. 2 60-175 mg / m² per week without interruption 2 20-150 mg / m² twice a week 2 , and 150-250 mg / m² twice a week 2 Examples include, but are not limited to, the following. The frequency of administration of mTOR inhibitor nanoparticle compositions (e.g., sirolimus / albumin nanoparticle compositions) may be adjusted throughout the course of treatment at the discretion of the prescribing physician.

[0078] In some embodiments, individuals are treated for at least one, two, three, four, five, six, seven, eight, nine, or ten treatment cycles.

[0079] The mTOR inhibitor nanoparticle compositions described herein (e.g., sirolimus / albumin nanoparticle compositions) allow for the injection of the mTOR inhibitor nanoparticle compositions into an individual over an injection time shorter than about 24 hours. For example, in some embodiments, the mTOR inhibitor nanoparticle compositions (e.g., sirolimus / albumin nanoparticle compositions) are administered over an injection period of less than 24 hours, 12 hours, 8 hours, 5 hours, 3 hours, 2 hours, 1 hour, 30 minutes, 20 minutes, or 10 minutes. In some embodiments, the mTOR inhibitor nanoparticle compositions (e.g., sirolimus / albumin nanoparticle compositions) are administered over an injection period of about 30 minutes.

[0080] In some embodiments, the exemplary dose of the mTOR inhibitor (in some embodiments, a limus-based drug, e.g., sirolimus) in the mTOR inhibitor nanoparticle composition is approximately 50 mg / m². 2 , 60 mg / m² 2 75 mg / m² 2 , 80 mg / m² 2 90 mg / m² 2 , 100 mg / m² 2 , 120 mg / m² 2 , 160 mg / m² 2 , 175 mg / m² 2 , 200 mg / m² 2 , 210 mg / m² 2 , 220 mg / m² 2 , 260 mg / m² 2 , and 300 mg / m² 2 Examples include, but are not limited to, any of the following. For example, the dose of an mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or its derivatives) in a nanoparticle composition is approximately 100-400 mg / m² when administered on a 3-week schedule. 2 or approximately 10-250 mg / m² if given on a weekly schedule. 2 It can be within the range.

[0081] In some embodiments, the dose of the mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus) is about 100 mg to about 400 mg, for example, about 100 mg, about 200 mg, about 300 mg, or about 400 mg. In some embodiments, the limus-based drug is administered at doses of about 100 mg per week, about 200 mg per week, about 300 mg per week, about 100 mg twice per week, or about 200 mg twice per week. In some embodiments, this is followed by a monthly maintenance dose (which may be the same as or different from the weekly dose).

[0082] In some embodiments, when a limus-based nanoparticle composition is administered intravenously, the dose of the mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus) in the nanoparticle composition can be in the range of about 30 mg to about 400 mg. The mTOR inhibitor nanoparticle compositions described herein (e.g., sirolimus / albumin nanoparticle compositions) allow for the injection of the mTOR inhibitor nanoparticle composition into an individual over an injection time shorter than about 24 hours. For example, in some embodiments, the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition) is administered over an injection period of about 24 hours, 12 hours, 8 hours, 5 hours, 3 hours, 2 hours, 1 hour, 30 minutes, 20 minutes, or less than 10 minutes. In some embodiments, the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition) is administered over an injection period of about 30 minutes to about 40 minutes.

[0083] The anti-PD-1 antibodies described herein (e.g., nivolumab) can be injected into an individual over an injection time shorter than approximately 24 hours. For example, in some embodiments, the anti-PD-1 antibody (e.g., nivolumab) is administered over an injection period of approximately 24 hours, 12 hours, 8 hours, 5 hours, 3 hours, 2 hours, 1 hour, 30 minutes, 20 minutes, or less than 10 minutes. In some embodiments, the anti-PD-1 antibody (e.g., nivolumab) is administered over an injection period of approximately 30 minutes.

[0084] In some embodiments, each dose contains both an mTOR inhibitor nanoparticle composition (e.g., a sirolimus / albumin nanoparticle composition) and an anti-PD-1 antibody delivered as a single dose, while in other embodiments, each dose contains either the mTOR inhibitor nanoparticle composition or an anti-PD-1 antibody delivered as a separate dose.

[0085] mTOR inhibitor nanoparticle compositions (e.g., sirolimus / albumin nanoparticle compositions) and anti-PD-1 antibodies, in their pure form or in suitable pharmaceutical compositions, can be administered by any of the accepted modes of administration or drugs known in the art. The compositions and / or drugs can be administered, for example, orally, nasally, parenterally (e.g., intravenously, intramuscularly, or subcutaneously), topically, transdermally, vaginally, intravesicularly, intracapsularly, or rectally. Dosage forms can be, for example, solid, semi-solid, lyophilized powder, or liquid dosage forms, such as tablets, pills, soft elastic or hard gelatin capsules, powders, solutions, suspensions, suppositories, aerosols, etc., preferably unit dosage forms suitable for simple administration of precise doses.

[0086] As described above, the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition) and the second therapeutic agent (e.g., anti-PD-1 antibody) can be administered in a single unit dose or in separate dosage forms. Therefore, the term “pharmaceutically acceptable combination” includes a combination of two drugs in either a single dosage form or separate dosage forms, i.e., the pharmaceutically acceptable carriers and excipients described throughout this application can be combined with the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition) and the second therapeutic agent (e.g., anti-PD-1 antibody) in a single unit dose, and can also be combined individually with the mTOR inhibitor nanoparticle composition and the second therapeutic agent (e.g., anti-PD-1 antibody) when these compounds are administered separately.

[0087] Auxiliaries and adjuvants may include, for example, preservatives, humectants, suspending agents, sweeteners, flavoring agents, fragrances, emulsifiers, and dispersing agents. Prevention of microbial action is generally provided by various antimicrobial and antifungal agents, such as parabens, chlorobutanol, phenol, and sorbic acid. Isotonic agents, such as sugars and sodium chloride, may also be included. Sustained absorption of injectable pharmaceutical forms can be achieved by the use of absorption-delaying agents, such as aluminum monostearate and gelatin. Auxiliaries may also include humectants, emulsifiers, pH buffers, and antioxidants, such as citric acid, sorbitan monolaurate, triethanolamine oleate, and butylated hydroxytoluene.

[0088] Solid dosage forms can be prepared using coatings and shells, such as enteric coatings and others well known in the art. These may contain sedatives and may be compositions that release the active compound(s) in a delayed manner in a specific portion of the intestinal tract. Examples of embedding compositions that can be used are polymeric substances and waxes. The active compound may also be in a microencapsulated form together with one or more of the excipients described above, where appropriate.

[0089] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solvents, suspensions, syrups, and elixirs. Such dosage forms are prepared, for example, by dissolving or dispersing an mTOR inhibitor nanoparticle composition described herein (e.g., sirolimus / albumin nanoparticle composition) or an anti-PD-1 antibody, or a pharmaceutically acceptable salt thereof, and any pharmaceutically acceptable adjuvant in a carrier such as water, physiological saline, aqueous dextrose, glycerol, or ethanol, a solubilizer and emulsifier such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, or dimethylformamide, an oil, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and sorbitan fatty acid esters, or mixtures thereof, thereby forming a solution or suspension.

[0090] In some embodiments, depending on the intended mode of administration, the pharmaceutically acceptable composition contains about 1% to about 99% by weight of the compound described herein or a pharmaceutically acceptable salt thereof, and 99% to 1% by weight of a pharmaceutically acceptable excipient. In one example, the composition contains about 5% to about 75% by weight of the compound described herein or a pharmaceutically acceptable salt thereof, with the remainder being a preferred pharmaceutically acceptable excipient.

[0091] Practical methods for preparing such dosage forms will be known or apparent to those skilled in the art. See, for example, Remington's Pharmaceutical Sciences, 18th Ed., (Mack Publishing Company, Easton, Pa., 1990).

[0092] mTOR inhibitor nanoparticle compositions (e.g., sirolimus / albumin nanoparticle compositions) can be administered to an individual (e.g., a human) via various routes, including intravenous, intraarterial, intraperitoneal, intrapulmonary, oral, inhalation, intravesicular, intramuscular, intratracheal, subcutaneous, intraocular, intrathecal, transmucosal, and transdermal. In some embodiments, a sustained-release formulation of the composition may be used. In some embodiments, the composition is administered intravenously. In some embodiments, the composition is administered intraportally. In some embodiments, the composition is administered intraarterially. In some embodiments, the composition is administered intraperitoneally.

[0093] Anti-PD-1 antibodies (e.g., nivolumab) can be administered to an individual (e.g., a human) via various routes, including intravenous, intra-arterial, intraperitoneal, intrapulmonary, oral, inhalation, intravesicular, intramuscular, intratracheal, subcutaneous, intraocular, intrathecal, transmucosal, and transdermal. In some embodiments, the anti-PD-1 antibody is administered intravenously.

[0094] B. Treatment of undifferentiated pleomorphic sarcoma In some embodiments, a method is provided for treating undifferentiated pleomorphic sarcoma in an individual requiring treatment, the method comprising administering to the individual (a) an effective amount of a composition comprising nanoparticles containing an mTOR inhibitor and albumin, and optionally (b) an effective amount of a second therapeutic agent (e.g., an anti-PD-1 antibody).

[0095] Undifferentiated pleomorphic sarcoma (UPS) is a highly malignant, often invasive soft tissue sarcoma. It typically presents as an asymptomatic, inconspicuous, rapidly growing skin or subcutaneous nodule, usually without surface skin abnormalities. The origin of UPS is likely mesenchymal stem cells. It can manifest as affecting bone, soft tissue, and retroperitoneum, among other areas, and can also metastasize to other organs. UPS can be diagnosed through histopathology of tumor specimens from core-needle techniques or biopsies. Markers can be used to diagnose UPS and include keratin, S100 protein, and / or SOX10, smooth muscle actin (SMA), and desmin. MDM2 and CDK4 may also be useful in distinguishing UPS from dedifferentiated liposarcoma. Undifferentiated pleomorphic sarcomas exhibit atypical pleomorphic spindle cells with abundant mitotic indices, and the tumor may show malformed, fascicular, or sheet-like structures within the fibrous stroma. Robles-Tenorio & Solis-Lesema, StatPearls, Undifferentiated Pleomorphic Sarcoma, 2022.

[0096] In some embodiments, undifferentiated pleomorphic sarcomas exhibit a loss of phosphatase tensin homolog (PTEN). In some embodiments, PTEN loss is due to loss-of-function mutations or epigenetic silencing. PTEN loss is described, for example, in Chang et al., Biomolecules, 9, 2019 and Vidotto et al., BrJ Cancer, 122, 2020, the contents of which, in whole, are incorporated herein by reference.

[0097] In some embodiments, undifferentiated pleomorphic sarcoma has a tuberous sclerosis complex 2 (TSC2) mutation. In some embodiments, the TSC2 mutation is a missense mutation, a nonsense mutation, a deletion, a splicing site mutation, an insertion, a substation, a rearrangement, or a frameshift, or a combination thereof. In some embodiments, the TSC2 mutation is an abnormal mTOR activation of TSC2. In some embodiments, the TSC2 mutation includes a single nucleotide variant (SNV). In some embodiments, the SNV includes a mutation selected from the group consisting of C1503T, C2743G, C5383T, C3755G, G760T, C3442T, G880A, T707C, A4949G, or a deletion of one or more amino acids at the positions 1405-1409, 1960-1970, 4999, 5002, 3521, 5208, 5238-5255. In some embodiments, the TSC2 mutation is a copy number variation of TSC2. In some embodiments, the TSC2 mutation is a loss-of-function mutation. In some embodiments, the TSC2 mutation results in an abnormal level of TSC2 expression. In some embodiments, the TSC2 mutation results in an abnormal level of activity of the protein encoded by TSC2. In some embodiments, the TSC2 mutation results in a loss of heterozygosity of TSC2. Mutational analysis of the TSC2 gene is known, e.g., Avgeris et al., Sci Rep, 7, 2017, the contents of which are incorporated herein by reference in whole.

[0098] In some embodiments, individuals are selected for treatment based on having PTEN loss. In some embodiments, individuals are selected for treatment based on a TSC2 mutation. In some embodiments, individuals are selected for treatment based on having both PTEN loss and a TSC2 mutation. In some embodiments, the method further includes selecting individuals based on having PTEN loss. In some embodiments, the method further includes selecting individuals based on having a TSC2 mutation. In some embodiments, the method further includes selecting individuals based on having both PTEN loss and a TSC2 mutation.

[0099] In some embodiments, a method is provided for treating undifferentiated pleomorphic sarcoma with PTEN loss in an individual (e.g., human), the method comprising administering to the individual (a) an effective amount of a composition comprising nanoparticles containing an mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or a derivative thereof) and albumin, and optionally (b) an effective amount of a second therapeutic agent (e.g., an anti-PD-1 antibody). In some embodiments, the method comprises administering to the individual (a) an effective amount of a composition comprising nanoparticles containing an mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or a derivative thereof) and albumin, wherein the mTOR inhibitor in the nanoparticles is contained within the nanoparticles that are associated with (e.g., coated) albumin, and (b) an effective amount of an anti-PD-1 antibody. In some embodiments, the method involves administering to an individual an effective amount of (a) an effective amount of a composition comprising nanoparticles containing an mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or a derivative thereof) and albumin, wherein the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less), and (b) an effective amount of an anti-PD-1 antibody. In some embodiments, the method comprises administering to an individual an effective amount of (a) an effective amount of a composition comprising nanoparticles containing an mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or a derivative thereof) and albumin, wherein the nanoparticles are associated with (e.g., coated with) the mTOR inhibitor, the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), and the weight ratio of albumin to mTOR inhibitor in the mTOR inhibitor nanoparticle composition is about 9:1 or less (e.g., about 9:1 or about 8:1); and (b) an effective amount of an anti-PD-1 antibody.In some embodiments, the mTOR inhibitor is a limus-based drug. In some embodiments, the mTOR inhibitor is sirolimus or a derivative thereof. In some embodiments, the mTOR inhibitor nanoparticle composition comprises nab-sirolimus. In some embodiments, the mTOR inhibitor nanoparticle composition is nab-sirolimus. In some embodiments, the anti-PD-1 antibody is selected from the group consisting of nivolumab, pembrolizumab, semiprimab, atezolizumab, dostallimab, durvalumab, and avelumab. In some embodiments, the anti-PD-1 antibody is nivolumab. In some embodiments, the nanoparticle composition and the anti-PD-1 antibody are administered sequentially. In some embodiments, the nanoparticle composition and the anti-PD-1 antibody are administered simultaneously. In some embodiments, the nanoparticle composition and the anti-PD-1 antibody are administered in parallel. In some embodiments, the mTOR inhibitor nanoparticle composition is administered for 2 weeks every 3 weeks, for example, on days 8 and 15 of a 21-day cycle. In some embodiments, the amount of mTOR inhibitor in the mTOR inhibitor nanoparticle composition is approximately 10 mg / m. 2 ~about 150mg / m 2 For example, approximately 100 mg / m² 2 In some embodiments, the anti-PD-1 antibody is administered every three weeks, for example, on day 1 of a 21-day cycle. In some embodiments, the anti-PD-1 antibody is administered for at least one cycle prior to the administration of the mTOR inhibitor nanoparticle composition. In some embodiments, the anti-PD-1 antibody is administered in an amount of about 1 mg / kg to about 10 mg / kg, for example, about 3 mg / kg.

[0100] In some embodiments, a method is provided for treating an undifferentiated pleomorphic sarcoma having a TSC2 mutation in an individual (e.g., a human), the method comprising administering to the individual (a) an effective amount of a composition comprising nanoparticles containing an mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or a derivative thereof) and albumin, and optionally (b) an effective amount of a second therapeutic agent (e.g., an anti-PD-1 antibody). In some embodiments, the method comprises administering to the individual (a) an effective amount of a composition comprising nanoparticles containing an mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or a derivative thereof) and albumin, wherein the mTOR inhibitor in the nanoparticles is contained within nanoparticles that are associated with albumin (e.g., coated), and (b) an effective amount of an anti-PD-1 antibody. In some embodiments, the method involves administering to an individual an effective amount of (a) an effective amount of a composition comprising nanoparticles containing an mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or a derivative thereof) and albumin, wherein the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less), and (b) an effective amount of an anti-PD-1 antibody. In some embodiments, the method comprises administering to an individual an effective amount of (a) an effective amount of a composition comprising nanoparticles containing an mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or a derivative thereof) and albumin, wherein the nanoparticles are associated with (e.g., coated with) the mTOR inhibitor, the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), and the weight ratio of albumin to mTOR inhibitor in the mTOR inhibitor nanoparticle composition is about 9:1 or less (e.g., about 9:1 or about 8:1); and (b) an effective amount of an anti-PD-1 antibody.In some embodiments, the mTOR inhibitor is a limus-based drug. In some embodiments, the mTOR inhibitor is sirolimus or a derivative thereof. In some embodiments, the mTOR inhibitor nanoparticle composition comprises nab-sirolimus. In some embodiments, the mTOR inhibitor nanoparticle composition is nab-sirolimus. In some embodiments, the anti-PD-1 antibody is selected from the group consisting of nivolumab, pembrolizumab, semiprimab, atezolizumab, dostallimab, durvalumab, and avelumab. In some embodiments, the anti-PD-1 antibody is nivolumab. In some embodiments, the nanoparticle composition and the anti-PD-1 antibody are administered sequentially. In some embodiments, the nanoparticle composition and the anti-PD-1 antibody are administered simultaneously. In some embodiments, the nanoparticle composition and the anti-PD-1 antibody are administered in parallel. In some embodiments, the mTOR inhibitor nanoparticle composition is administered for 2 weeks every 3 weeks, for example, on days 8 and 15 of a 21-day cycle. In some embodiments, the amount of mTOR inhibitor in the mTOR inhibitor nanoparticle composition is approximately 10 mg / m. 2 ~about 150mg / m 2 For example, approximately 100 mg / m² 2 In some embodiments, the anti-PD-1 antibody is administered every three weeks, for example, on day 1 of a 21-day cycle. In some embodiments, the anti-PD-1 antibody is administered for at least one cycle prior to the administration of the mTOR inhibitor nanoparticle composition. In some embodiments, the anti-PD-1 antibody is administered in an amount of about 1 mg / kg to about 10 mg / kg, for example, about 3 mg / kg.

[0101] In some embodiments, a method is provided for treating undifferentiated pleomorphic sarcoma having PTEN loss and TSC2 mutation in an individual (e.g., human), the method comprising administering to the individual (a) an effective amount of a composition comprising nanoparticles containing an mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or a derivative thereof) and albumin, and optionally (b) an effective amount of a second therapeutic agent (e.g., an anti-PD-1 antibody). In some embodiments, the method comprises administering to the individual (a) an effective amount of a composition comprising nanoparticles containing an mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or a derivative thereof) and albumin, wherein the mTOR inhibitor in the nanoparticles is contained within nanoparticles that are associated with albumin (e.g., coated), and (b) an effective amount of an anti-PD-1 antibody. In some embodiments, the method involves administering to an individual an effective amount of (a) an effective amount of a composition comprising nanoparticles containing an mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or a derivative thereof) and albumin, wherein the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less), and (b) an effective amount of an anti-PD-1 antibody. In some embodiments, the method comprises administering to an individual an effective amount of (a) an effective amount of a composition comprising nanoparticles containing an mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or a derivative thereof) and albumin, wherein the nanoparticles are associated with (e.g., coated with) the mTOR inhibitor, the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), and the weight ratio of albumin to mTOR inhibitor in the mTOR inhibitor nanoparticle composition is about 9:1 or less (e.g., about 9:1 or about 8:1); and (b) an effective amount of an anti-PD-1 antibody.In some embodiments, the mTOR inhibitor is a limus-based drug. In some embodiments, the mTOR inhibitor is sirolimus or a derivative thereof. In some embodiments, the mTOR inhibitor nanoparticle composition comprises nab-sirolimus. In some embodiments, the mTOR inhibitor nanoparticle composition is nab-sirolimus. In some embodiments, the anti-PD-1 antibody is selected from the group consisting of nivolumab, pembrolizumab, semiprimab, atezolizumab, dostallimab, durvalumab, and avelumab. In some embodiments, the anti-PD-1 antibody is nivolumab. In some embodiments, the nanoparticle composition and the anti-PD-1 antibody are administered sequentially. In some embodiments, the nanoparticle composition and the anti-PD-1 antibody are administered simultaneously. In some embodiments, the nanoparticle composition and the anti-PD-1 antibody are administered in parallel. In some embodiments, the mTOR inhibitor nanoparticle composition is administered for 2 weeks every 3 weeks, for example, on days 8 and 15 of a 21-day cycle. In some embodiments, the amount of mTOR inhibitor in the mTOR inhibitor nanoparticle composition is approximately 10 mg / m. 2 ~about 150mg / m 2 For example, approximately 100 mg / m² 2 In some embodiments, the anti-PD-1 antibody is administered every three weeks, for example, on day 1 of a 21-day cycle. In some embodiments, the anti-PD-1 antibody is administered for at least one cycle prior to the administration of the mTOR inhibitor nanoparticle composition. In some embodiments, the anti-PD-1 antibody is administered in an amount of about 1 mg / kg to about 10 mg / kg, for example, about 3 mg / kg.

[0102] C. Treatment of leiomyosarcoma In some embodiments, this specification provides a method for treating leiomyosarcoma in an individual requiring treatment, the method comprising administering to the individual (a) an effective amount of a composition comprising nanoparticles containing an mTOR inhibitor and albumin, and optionally (b) an effective amount of a second therapeutic agent (e.g., an anti-PD-1 antibody).

[0103] Leiomyosarcoma is a subtype of soft tissue sarcoma that is found in most parts of the human body, with common locations including the abdomen, retroperitoneum, larger blood vessels, and uterus. Leiomyosarcoma is a malignant mesenchymal tumor composed of cells exhibiting distinct features of the smooth muscle lineage and can be classified as somatic soft tissue leiomyosarcoma, cutaneous leiomyosarcoma, or angioleiomyosarcoma. Diagnosis can be made from tumor specimens such as those obtained from core needle biopsy. Leiomyosarcoma generally exhibits high cytoplasmic regions arranged in fibrous bundles, and malignant cells are characterized by cytoplasm ranging from rich pink to deep red with hematoxylin and eosin (H&E) staining, and have nuclei located in the center of the embryonic shape. Such distinguishing features are lost in dedifferentiated tumors. EL-Naggar et al., Cancer Genomics, Chapter 22, 2014.

[0104] In some embodiments, the leiomyosarcoma is an estrogen receptor-positive leiomyosarcoma.

[0105] In some embodiments, individuals are selected for treatment based on having estrogen receptor-positive leiomyosarcoma. In some embodiments, the method further includes selecting individuals based on having estrogen receptor-positive leiomyosarcoma.

[0106] In some embodiments, a method is provided for treating estrogen receptor site leiomyosarcoma in an individual (e.g., human), the method comprising administering to the individual (a) an effective amount of a composition comprising nanoparticles containing an mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or a derivative thereof) and albumin, and (b) an effective amount of an anti-PD-1 antibody. In some embodiments, the method comprises administering to the individual (a) an effective amount of a composition comprising nanoparticles containing an mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or a derivative thereof) and albumin, wherein the mTOR inhibitor in the nanoparticles is contained within the nanoparticles that are associated with (e.g., coated) albumin, and (b) an effective amount of an anti-PD-1 antibody. In some embodiments, the method involves administering to an individual an effective amount of (a) an effective amount of a composition comprising nanoparticles containing an mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or a derivative thereof) and albumin, wherein the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less), and (b) an effective amount of an anti-PD-1 antibody. In some embodiments, the method comprises administering to an individual an effective amount of (a) an effective amount of a composition comprising nanoparticles containing an mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or a derivative thereof) and albumin, wherein the nanoparticles are associated with (e.g., coated with) the mTOR inhibitor, the nanoparticles have an average particle size of about 150 nm or less (e.g., about 120 nm or less, e.g., about 100 nm), and the weight ratio of albumin to mTOR inhibitor in the mTOR inhibitor nanoparticle composition is about 9:1 or less (e.g., about 9:1 or about 8:1); and (b) an effective amount of an anti-PD-1 antibody.In some embodiments, the mTOR inhibitor is a limus-based drug. In some embodiments, the mTOR inhibitor is sirolimus or a derivative thereof. In some embodiments, the mTOR inhibitor nanoparticle composition comprises nab-sirolimus. In some embodiments, the mTOR inhibitor nanoparticle composition is nab-sirolimus. In some embodiments, the anti-PD-1 antibody is selected from the group consisting of nivolumab, pembrolizumab, semiprimab, atezolizumab, dostallimab, durvalumab, and avelumab. In some embodiments, the anti-PD-1 antibody is nivolumab. In some embodiments, the nanoparticle composition and the anti-PD-1 antibody are administered sequentially. In some embodiments, the nanoparticle composition and the anti-PD-1 antibody are administered simultaneously. In some embodiments, the nanoparticle composition and the anti-PD-1 antibody are administered in parallel. In some embodiments, the mTOR inhibitor nanoparticle composition is administered for 2 weeks every 3 weeks, for example, on days 8 and 15 of a 21-day cycle. In some embodiments, the amount of mTOR inhibitor in the mTOR inhibitor nanoparticle composition is approximately 10 mg / m. 2 ~about 150mg / m 2 For example, approximately 100 mg / m² 2 In some embodiments, the anti-PD-1 antibody is administered every three weeks, for example, on day 1 of a 21-day cycle. In some embodiments, the anti-PD-1 antibody is administered for at least one cycle prior to the administration of the mTOR inhibitor nanoparticle composition. In some embodiments, the anti-PD-1 antibody is administered in an amount of about 1 mg / kg to about 10 mg / kg, for example, about 3 mg / kg.

[0107] D. Treatment of other cancers In some embodiments, a method is provided for treating cancer in an individual in need of treatment, the cancer being selected from the group consisting of colorectal cancers such as ascending colon adenocarcinoma, cervical squamous cell carcinoma, chondrosarcoma, chordoma, clear cell sarcoma, metastatic colorectal cancer, osteosarcoma including fibrous round cell tumor, fibrous small cell tumor, Ewing's sarcoma, myxoid liposarcoma, osteogenic osteosarcoma, pleomorphic spindle cell sarcoma, serous carcinoma of the endometrium, or synovial sarcoma, and the method comprising administering to the individual (a) an effective amount of a composition comprising nanoparticles containing an mTOR inhibitor and albumin, and optionally (b) an effective amount of a second therapeutic agent (e.g., an anti-PD-1 antibody).

[0108] In some embodiments, the cancer is chondrosarcoma, and the individual has an abnormality in one or more genes (e.g., two or more, e.g., three or more) selected from the group consisting of TP53 p.Y220C, TUBB3, and AXL. In some embodiments, the cancer is chondrosarcoma, and the individual has an abnormality in one or more genes (e.g., two or more, e.g., three or more) selected from the group consisting of BRCA2-cleaved intron 7, EZH2-cleaved intron 19, MLL2 R5048H, RB1 exon 3-9 loss, and TP53 R110del. In some embodiments, the cancer is chondrosarcoma, and the individual has an abnormality in IDH1. In some embodiments, the cancer is chondrosarcoma, and the individual has an abnormality in IDH1. In some embodiments, the cancer is chondrosarcoma, and the individual has one or more abnormalities (e.g., two or more, e.g., three or more) selected from the group consisting of tumor gene mutational load (TMB-low), mismatch repair (MMR)-normal, PD-L1-negative, MLH1-positive, MLH2-positive, MLH6-positive, and PMS2-positive.

[0109] In some embodiments, the cancer is osteosarcoma, and the individual has the C17orf39 mutation and is MUTYH-positive. In some embodiments, the cancer is osteosarcoma, and the individual has abnormalities in DPB1*04:01 and / or DPBI*04:02, and / or is DPw4-positive. In some embodiments, the cancer is chondrosarcoma, and the individual has abnormalities in one or more genes (e.g., two or more, e.g., three or more) selected from the group consisting of BRCA2, FGFR1, KDM5A, MAF, MYO18A, PCLO, RELN, and SF3B1, and / or the individual is RB1-positive and TP53-positive. In some embodiments, the cancer is osteosarcoma, and the individual is microsatellite (MSI)-stable, MMR-normal, TMB-intermediate, and / or TP53-positive. In some embodiments, the cancer is osteosarcoma, and the individual is TMB-intermediate, PD-L1-negative, MLH1-positive, MLH2-positive, MLH6-positive, PMS2-positive, and / or has an abnormality in TP53.

[0110] In some embodiments, the cancer is Ewing sarcoma, and the individual has an abnormality in EWSR1 (e.g., a gene fusion containing at least a portion of the EWSR1 gene) and / or KMT2D-normal. In some embodiments, the cancer is Ewing sarcoma, and the individual has an abnormality in EWSR1 (e.g., a gene fusion containing at least a portion of the EWSR1 gene, e.g., an EWSR1-FLI1 fusion) and / or tumor exonic gene mutational load-high, MLH1-positive, MLH2-positive, MLH6-positive and / or PMS2-positive. In some embodiments, the cancer is Ewing sarcoma, and the individual has an abnormality in EWSR1 (e.g., a gene fusion containing at least a portion of the EWSR1 gene) and / or EZH2 (e.g., a pathogenic variant of EZH2) and / or MLH1-positive, MLH2-positive, MLH6-positive and / or PMS2-positive.

[0111] In some embodiments, the cancer is synovial sarcoma, and the individual is NY-ESO-1 positive (e.g., 99% of tumor cells are NY-ESO-1 positive).

[0112] In some embodiments, the cancer is a fibrous round cell tumor, and the individual has an abnormality in EWSR1 (e.g., a gene fusion containing at least a portion of the EWSR1 gene, e.g., an EWSR1-WT1 fusion) and / or TMB-low.

[0113] In some embodiments, the cancer is a clear cell sarcoma, and the individual is EWSR-positive.

[0114] In some embodiments, the cancer is a chordoma, and the individual has an abnormality in one (e.g., two or more, e.g., three or more) gene selected from the group consisting of ARAF, ARID1A, ASXL1, BAP1, DNMT3A, FANCE, FGFR4, FLT3, FLT4, JAK3, KDM5C, NF1, PTCH, TERT, and TP53, and / or is MSI-stable, TMB-low, TSC2-positive, and / or INI-1-positive.

[0115] In some embodiments, the cancer is a serous carcinoma of the endometrium, and the individual has an abnormality in PIK3CA and / or is TP53-positive.

[0116] In some embodiments, the cancer is metastatic colorectal cancer, and the individual has abnormalities in CRKL (e.g., amplification of the CRKL locus) and / or TP53 (e.g., a pathogenic variant of TP53, e.g., p.T253P), and / or is MSI-high, MMR-deficient (dMMR), PTEN-positive, MLH1-positive, MLH2-positive, MLH6-positive, and / or PMS2-positive.

[0117] In some embodiments, the cancer is cervical squamous cell carcinoma, the individual has abnormalities in ERBB2 and / or AKT2 (e.g., amplification of the AKT2 locus), and / or is PD-L1-positive and / or TMB-intermediate.

[0118] In some embodiments, the cancer is osteosarcoma of osteogenic origin, and the individual has an abnormality in one (e.g., two or more, e.g., three or more) of the genes selected from the group consisting of BRCA2, FGFR1, KDM5A, MAF, MYO18A, PCLO, RELN, and SF3B1, and is / or RB1-positive and / or TP53-positive.

[0119] In some embodiments, the cancer is an ascending colon adenocarcinoma, and the individual has an abnormality in one or more genes (e.g., two or more, e.g., three or more) selected from the group consisting of KRAS, NRAS, APC, BRCA, PIK3CA, SMAD4, and TP53, and / or is a TMB intermediate.

[0120] In some embodiments, the cancer is pleomorphic spindle cell sarcoma, and the individual has abnormalities in one or more genes (e.g., two or more, e.g., three or more) selected from the group consisting of APC, ATRX, BRCA2, FBXW7, FLT1, GNA13, IRS2, JAK3, KMT2A, MED12, MLL2, PRKAR1A, and SOX9, and / or is microsatellite stability (MSS), TMB-low, CCND3-positive, RB1-positive, VEGFR-positive, PDL-1-positive, and / or TSC2-positive.

[0121] In some embodiments, as used herein, an individual being "-positive" for a biomarker, for example, "RB1-positive," means that the individual has detectable expression of the biomarker, such that it can be detected via standard biomarker measurement techniques including immunohistochemistry, mass spectrometry, PCR, or sequencing. In some embodiments, biomarker expression is detectable if the normalized reads per kilobase million (RPKM), normalized fragments per kilobase million (FPKM), or normalized transcripts per kilobase million (TPM) of the biomarker is 1 or greater. In some embodiments, as used herein, an individual being "-negative" for a biomarker, for example, "PD-L1-negative," means that the individual does not have detectable expression of the biomarker, such that it can be detected via standard biomarker measurement techniques including immunohistochemistry, mass spectrometry, PCR, or sequencing.

[0122] E. Biomarkers In certain embodiments of the methods provided herein, the individuals are AKT1, AKT2, APC, ARAF, ARID1A, ASXL1, ATRX, AXL, BAP1, BRCA, BRCA2, C17orf39, CCND3, CRKL, DNMT3A, ERBB2, EWSR1, EZH2, FANCE, FBXW7, FGFR1, FGFR4, FLI1, FLT1, FLT3, FLT4, GNA13, HLA-A2, HLA-DPB1, IDH1, INI-1, IRS2, JAK3, KDM5A, KDM5C, KMT2 Individuals are selected for the treatment described herein based on having an abnormality in one or more genes (e.g., two or more, e.g., three or more) selected from A, KMT2D, KRAS, MAF, MED12, MLL2, MUTYH, MYO18A, NF1, NRAS, NTRK1 / 2 / 3, NY-ESO-1, PCLO, PIK3CA, PRKAR1A, PTCH, PD-L1, PTEN, RB1, RELN, SF3B1, SMAD4, SOX9, TERT, TP53, TSC2, TUBB3, VEGFA, WT1. In some embodiments, individuals are selected for treatment based on an abnormality in PTEN and / or TSC2. In some embodiments, the abnormality in PTEN is PTEN loss. In some embodiments, the abnormality in TSC2 is a TSC2 mutation. In some embodiments, the abnormality in BRCA2 is a cleavage in BRCA2 intron 7. In some embodiments, the abnormality in EZH2 is a cleavage in intron 19. In some embodiments, the abnormality in EWSR1 is an EWSR1 fusion. In some embodiments, the EWSR1 fusion is an EWSR1-FLI1 fusion. In some embodiments, the abnormality in EWSR1 is an EWSR1 gene rearrangement. In some embodiments, the abnormality in FLI1 is an FLI1 fusion. In some embodiments, the FLI1 fusion is an EWSR1-FLI1 fusion. In some embodiments, the abnormality in MLL2 is an R5048H mutation. In some embodiments, the abnormality in RB1 is the loss of RB1 exons 3-9. In some embodiments, the abnormality in TP53 is TP53 p.Y220C. In some embodiments, the abnormality in TP53 is TP53 p.T253P.In some embodiments, the abnormality in TP53 is TP53 R110del. In some embodiments, the abnormality in AKT2 is gene amplification of AKT2. In some embodiments, the abnormality in CRKL is gene amplification of CRKL. In some embodiments, the abnormality in HLA-DPB1 is DPB1*04:01 and / or DPBI*04:02. In some embodiments, individuals are selected for treatment based on the absence of fusions in NTRK1 / 2 / 3.

[0123] In some embodiments, individuals are selected for treatment based on having detectable expression of a biomarker, for example, detectable expression via standard biomarker measurement techniques including immunohistochemistry, mass spectrometry, PCR, or sequencing. In some embodiments, biomarker expression is detectable if the normalized reads per kilobase million (RPKM), normalized fragments per kilobase million (FPKM), or normalized transcripts per kilobase million (TPM) of the biomarker is 1 or greater. In some embodiments, individuals are selected for treatment based on being DPw4-positive. In some embodiments, individuals are selected for treatment based on being INI-1-positive. In some embodiments, individuals are selected for treatment based on being NYESO-positive and HLA-A2-positive. In some embodiments, individuals are selected for treatment based on being PTEN-positive. In some embodiments, individuals are selected for treatment based on not having detectable expression of a biomarker. In some embodiments, individuals are selected for treatment based on being PD-L1-negative. In some embodiments, individuals are selected for treatment based on having a mutant state or genotype. In some embodiments, individuals are selected for treatment based on having a low tumor mutational load (TMB). In some embodiments, individuals are selected for treatment based on having a TMB intermediate. In some embodiments, individuals are selected for treatment based on having a high tumor exonic mutational load. In some embodiments, individuals are selected for treatment based on having microsatellite stability (MSS). In some embodiments, individuals are selected for treatment based on having high microsatellite instability and / or mismatch repair (MMR) deficiency.

[0124] In some embodiments, tumor mutational load (TMB) is the number of somatic (non-hereditary) mutations per megabase (Mb) of the genome. TMB can be used as a metric to predict outcomes such as the success of cancer treatment and survival rate. TMB can be measured using high-throughput sequencing techniques such as NGS. Different cancers may have variations in TMB levels. TMB levels can be classified as high, medium, and low based on the number of mutations / Mb. For example, in some embodiments, TMB levels can be set as (i) low - 1 to 5 mutations / Mb, (ii) medium - 6 to 15 mutations / Mb, and (iii) high - 16 or more mutations / Mb.

[0125] Mismatch repair (MMR) is a system for correcting base pair mismatches, insertions, or deletion errors during DNA replication and is crucial for maintaining genomic stability. MMR defects are associated with genomic instability and an increase in sporadic mutations in microsatellite regions. MMR status can be assessed by immunohistochemical staining to detect the presence or absence of MMR-regulated proteins MLH1, MSH2, MSH6, and PMS2. MMR status can be classified as normal MMR (pMMR) or deficient MMR (dMMR). pMMR status is associated with a functional MMR pathway, while dMMR is associated with the absence of one or more MMR proteins.

[0126] Microsatellite instability (MSI) is a phenotype that occurs when there are defects in DNA mismatch repair. MSI is a marker of cancer MMR status. MSI results in the insertion or deletion of repetitive sequences in the genome. MSI status is tested using PCR-based assays to determine instability using five markers: BAT-25, BAT-26, D2S123, D5S346, and D17S250. MSI status can be subdivided into high microsatellite instability (MSI-H: two or more markers are unstable), low microsatellite instability (MSI-L: one marker is unstable), and microsatellite stability (MSS: no markers show instability).

[0127] Where used herein, “based on” includes evaluating, determining, or measuring the characteristics of an individual as described herein (and preferably selecting an individual suitable for treatment). Where an abnormal condition is “used as a basis” for a method of selecting, evaluating, measuring, or determining a treatment as described herein, abnormalities in one or more genes are determined before and / or during treatment, and the resulting condition (including the presence, absence, expression level, activity level, and / or phosphorylation level of the abnormality) is used by a clinician in evaluating any of the following: (a) the likelihood or prospect of an individual being suitable or likely to receive the first treatment; (b) the likelihood or prospect of an individual being unsuitable or likely to receive the first treatment; (c) responsiveness to treatment; (d) the likelihood or prospect of an individual being suitable or likely to receive continued treatment; (e) the likelihood or prospect of an individual being unsuitable or likely to receive continued treatment; (f) dose adjustment; or (g) predicting potential clinical benefit. Where used herein, if an individual is selected for treatment "on the basis" of having an abnormality in one or more gene abnormalities (including the presence, absence, expression level, activity level, and / or phosphorylation level of the abnormality), the individual may be selected for treatment on two or more basis, and the basis for selection for treatment may include further criteria.

[0128] F. Techniques for determining cancer and / or individual conditions In certain embodiments of the methods provided herein, undifferentiated pleomorphic sarcoma is characterized by having PTEN loss and / or a TSC2 mutation. In certain embodiments of the methods provided herein, leiomyosarcoma is characterized by being estrogen receptor-positive leiomyosarcoma. In some embodiments, the methods provided herein include steps for evaluating and / or determining the cancerous state, such as undifferentiated pleomorphic sarcoma and / or leiomyosarcoma, in an individual. In some embodiments, the method includes obtaining a sample of cancer, such as undifferentiated pleomorphic sarcoma and / or leiomyosarcoma, from an individual. In some embodiments, the method includes determining the presence, absence, or level of a biomarker, such as the presence of PTEN loss, by means of, for example, protein expression, sequencing, or activity assays. In some embodiments, the method includes determining the presence of a TSC2 mutation, by means of, for example, protein expression, sequencing, or activity assays. In some embodiments, the method includes determining the presence, absence, or level of an estrogen receptor, by means of, for example, protein expression, sequencing, or activity assays.

[0129] For example, cancer and / or the state of the individual in undifferentiated pleomorphic sarcoma and / or leiomyosarcoma can be evaluated or determined by analyzing samples derived from the individual. The evaluation may be based on fresh tissue samples or archived tissue samples. Suitable samples include, but are not limited to, tumor tissue, normal tissue adjacent to tumor tissue, normal tissue distal to tumor tissue, or peripheral blood lymphocytes. In some embodiments, the sample is tumor tissue. In some embodiments, the sample is a biopsy containing tumor cells, e.g., microneedle aspiration of solid tumor cells or tumor cells obtained by laparoscopy. In some embodiments, the biopsy cells are centrifuged into a pellet, fixed, and embedded in paraffin before analysis. In some embodiments, these biopsy cells are rapidly frozen before analysis. In some embodiments, the sample is a plasma sample.

[0130] In some embodiments, the sample contains circulating metastatic cancer cells. In some embodiments, the sample is obtained by sorting circulating tumor cells (CTCs) from blood. In some further embodiments, the CTCs detach from the primary tumor and circulate in the body fluids. In some further embodiments, the CTCs detach from the primary tumor and circulate in the bloodstream. In some embodiments, the CTCs are indicators of metastasis.

[0131] In some embodiments, the sample is mixed with an antibody that recognizes a molecule (e.g., a protein) or a fragment thereof. In some embodiments, the sample is mixed with a nucleic acid that recognizes a nucleic acid (e.g., DNA or RNA) or a fragment thereof. In some embodiments, the sample is used for sequencing analysis, e.g., next-generation DNA, RNA, and / or exome sequencing analysis.

[0132] The sample may be evaluated before the start of treatment, at any point during the procedure, and / or at the end of the procedure.

[0133] III. Composition containing nanoparticles containing an mTOR inhibitor The mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising an mTOR inhibitor (e.g., a limus-based drug, e.g., rapamycin or its derivatives) and albumin (e.g., human serum albumin) (in various embodiments, essentially consisting of these or comprising them). Nanoparticles of poorly water-soluble drugs (e.g., macrolides) are disclosed, for example, in U.S. Patents Nos. 5,916,596, 6,506,405, 6,749,868, 6,537,579, 7,820,788, 8,911,786, and 11,497,737, and also in U.S. Patent Applications Nos. 2006 / 0263434 and 2007 / 0082838, PCT Patent Application No. W008 / 137148, and U.S. Patent Application No. 62 / 927,047, each of which is incorporated herein by reference as a whole.

[0134] In some embodiments, the composition includes nanoparticles having an average or intermediate diameter of about 1000 nanometers (nm) or less, for example, about 900, 800, 700, 600, 500, 400, 300, 200, and 100 nm or less. In some embodiments, the average or intermediate diameter of the nanoparticles is about 200 nm or less. In some embodiments, the average or intermediate diameter of the nanoparticles is about 150 nm or less. In some embodiments, the average or intermediate diameter of the nanoparticles is about 100 nm or less. In some embodiments, the average or intermediate diameter of the nanoparticles is about 10 to about 400 nm. In some embodiments, the average or intermediate diameter of the nanoparticles is about 10 to about 150 nm. In some embodiments, the average or intermediate diameter of the nanoparticles is about 40 nm to about 120 nm. In some embodiments, the average or intermediate diameter of the nanoparticles is about 50 nm or more. In some embodiments, the nanoparticles are sterile and filterable.

[0135] Methods for determining average particle size are well known in the art, and for example, dynamic light scattering (DLS) has been commonly used to determine the size of submicrometer-sized particles. See the international standard ISO 22412 Particle size analysis - dynamic light scattering, International Organization for Standardization (ISO) 2008 and definitions of general terms for dynamic light scattering (Malvern Instruments Limited, 2011). In some embodiments, particle size is measured as the volume-weighted average particle size (Dv50) of nanoparticles in a composition.

[0136] In some embodiments, the nanoparticles contain an mTOR inhibitor associated with albumin. In some embodiments, the nanoparticles contain an mTOR inhibitor coated with albumin.

[0137] In some embodiments, albumin has sulfhydryl groups that can form disulfide bonds. In some embodiments, at least about 5% of the albumin in the nanoparticle portion of the composition (e.g., including at least about 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%) is crosslinked (e.g., crosslinked via one or more disulfide bonds).

[0138] In some embodiments, nanoparticles containing an mTOR inhibitor (e.g., a limus-based drug, e.g., rapamycin or a derivative thereof) associate with (e.g., are coated with) albumin (e.g., human albumin or human serum albumin). In some embodiments, the composition contains an mTOR inhibitor (e.g., a limus-based drug, e.g., rapamycin or a derivative thereof) in both nanoparticle and non-nanoparticle forms (e.g., in solution form or in the form of a soluble albumin / nanoparticle complex), where at least about 50%, 60%, 70%, 80%, 90%, 95%, or 99% of the mTOR inhibitor in the composition is in nanoparticle form. In some embodiments, the mTOR inhibitor (e.g., a limus-based drug, e.g., rapamycin or a derivative thereof) in the nanoparticles constitutes about 50%, 60%, 70%, 80%, 90%, 95%, or 99% by weight of the nanoparticles. In some embodiments, the nanoparticles have a nonpolymer matrix. In some embodiments, the nanoparticles comprise a core of an mTOR inhibitor (e.g., a limousine drug, e.g., rapamycin or a derivative thereof) that is substantially free of polymer material (e.g., a polymer matrix).

[0139] In some embodiments, the composition comprises albumin in both the nanoparticle portion and the non-nanoparticle portion of the composition, with at least about 50%, 60%, 70%, 80%, 90%, 95%, or 99% of the albumin in the composition being in the non-nanoparticle portion of the composition.

[0140] In some embodiments, the weight ratio of albumin to the mTOR inhibitor (e.g., a limus-based drug, e.g., rapamycin or its derivatives) in the mTOR inhibitor nanoparticle composition is such that a sufficient amount of the mTOR inhibitor binds to or is transported by the cell. The weight ratio of albumin to mTOR inhibitors (e.g., limus-based drugs, e.g., rapamycin or its derivatives) must be optimized for different albumin-mTOR inhibitor combinations, but generally, the weight ratio (w / w) of albumin to mTOR inhibitors (e.g., limus-based drugs, e.g., rapamycin or its derivatives) is approximately 0.01:1 to 100:1, approximately 0.02:1 to 50:1, approximately 0.05:1 to 20:1, approximately 0.1:1 to 20:1, approximately 1:1 to 18:1, approximately 2:1 to 15:1, approximately 3:1 to 12:1, approximately 4:1 to 10:1, approximately 5:1 to 9:1, or approximately 9:1. In some embodiments, the weight ratio of albumin to an mTOR inhibitor (e.g., a limus-based drug, e.g., rapamycin or its derivatives) is one of the following: approximately 18:1 or less, 15:1 or less, 14:1 or less, 13:1 or less, 12:1 or less, 11:1 or less, 10:1 or less, 9:1 or less, 8:1 or less, 7:1 or less, 6:1 or less, 5:1 or less, 4:1 or less, and 3:1 or less. In some embodiments, the weight ratio of albumin (e.g., human albumin or human serum albumin) to mTOR inhibitor (e.g., limus-based drugs, e.g., rapamycin or its derivatives) in the composition is one of approximately 1:1 to approximately 18:1, approximately 1:1 to approximately 15:1, approximately 1:1 to approximately 12:1, approximately 1:1 to approximately 10:1, approximately 1:1 to approximately 9:1, approximately 1:1 to approximately 8:1, approximately 1:1 to approximately 7:1, approximately 1:1 to approximately 6:1, approximately 1:1 to approximately 5:1, approximately 1:1 to approximately 4:1, approximately 1:1 to approximately 3:1, approximately 1:1 to approximately 2:1, or approximately 1:1 to approximately 1:1.

[0141] In some embodiments, the composition comprises nanoparticles containing an mTOR inhibitor and albumin, with a weight ratio of albumin to mTOR inhibitor in the composition being about 0.01:1 to about 100:1. In some embodiments, the composition comprises nanoparticles containing an mTOR inhibitor (e.g., rapamycin) and albumin, with a weight ratio of albumin to mTOR inhibitor (e.g., rapamycin) in the composition being about 18:1 or less (including, for example, any of about 1:1 to about 18:1, about 2:1 to about 15:1, about 3:1 to about 12:1, about 4:1 to about 10:1, about 5:1 to about 9:1, and about 9:1). In some embodiments, the composition comprises nanoparticles containing rapamycin or a derivative thereof, and albumin, wherein the weight ratio of albumin to rapamycin or a derivative thereof in the composition is about 18:1 or less (e.g., including any of about 1:1 to about 18:1, about 2:1 to about 15:1, about 3:1 to about 12:1, about 4:1 to about 10:1, about 5:1 to about 9:1, and about 9:1). In some embodiments, the mTOR inhibitor (e.g., rapamycin) is coated with albumin.

[0142] In some embodiments, the mTOR inhibitor nanoparticle composition (e.g., rapamycin / albumin nanoparticle composition) includes one or more of the above-described features.

[0143] The nanoparticles described herein may be present in a dry formulation (e.g., a lyophilized composition) or suspended in a biocompatible medium. Suitable biocompatible mediums include, but are not limited to, water, buffered aqueous media, physiological saline, buffered physiological saline, optionally an amino acid buffer solution, optionally a protein buffer solution, optionally a sugar buffer solution, optionally a vitamin buffer solution, optionally a synthetic polymer buffer solution, and lipid-containing emulsions.

[0144] In some embodiments, the pharmaceutically acceptable carrier includes albumin (e.g., human albumin or human serum albumin). The albumin may be of natural origin or may be synthetically prepared. In some embodiments, the albumin is human albumin or human serum albumin. In some embodiments, the albumin is recombinant albumin.

[0145] Human serum albumin (HSA) is M r HSA is a highly soluble, globular protein with a 65K content, consisting of 585 amino acids. HSA is the most abundant protein in plasma, accounting for 70-80% of the colloidal osmotic pressure of human plasma. The amino acid sequence of HSA contains a total of 17 disulfide crosslinks, one free thiol (Cys 34), and one tryptophan (Trp 214). Intravenous use of HSA solution is indicated for the prevention and treatment of hypovolemic shock (see, e.g., Tullis, JAMA, 237:355-360, 460-463, (1977) and Houser et al., Surgery, Gynecology and Obstetrics, 150:811-816 (1980)), and in combination with exchange transfusion in the treatment of neonatal hyperbilirubinemia (see, e.g., Finlayson, Seminars in Thrombosis and Hemostasis, 6, 85-120, (1980)). Other albumins, such as bovine serum albumin, are being considered. The use of such non-human albumins is appropriate in the context of the use of these compositions in non-human mammals, such as in veterinary medicine (including in the contexts of domestic pets and agriculture). Human serum albumin (HSA) has multiple hydrophobic binding sites (a total of eight for fatty acids, which are the endogenous ligands of HSA), and binds to a diverse set of drugs, particularly neutral and negatively charged hydrophobic compounds (Goodman et al., The Pharmacological Basis of Therapeutics, 9). th(ed, McGraw-Hill New York (1996)). Two high-affinity binding sites have been proposed in subdomains IIA and IIIA of HSA, which are highly elongated hydrophobic pockets with charged lysine and arginine residues near the surface that function as attachment sites for polar ligand features (e.g., Fehske et al., Biochem. Pharmcol., 30, 687-92 (198a), Vorum, Dan. Med. Bull., 46, 379-99 (1999), Kragh-Hansen, Dan. Med. Bull., 1441, 131-40 (1990), Curry et al., Nat. Struct. Biol., 5, 827-35 (1998), Sugio et al., Protein. Eng., 12, 439-46 (1999), He et al. See al., Nature, 358, 209-15 (199b), and Carter et al., Adv. Protein. Chem., 45, 153-203 (1994). Rapamycin and propofol have been shown to bind to HSA (see, for example, Paal et al., Eur. J. Biochem., 268(7), 2187-91 (200a), Purcell et al., Biochem. Biophys. Acta, 1478(a), 61-8 (2000), Altmayer et al., Arzneimittelforschung, 45, 1053-6 (1995), and Garrido et al., Rev. Esp. Anestetiol. Reanim., 41, 308-12 (1994)). Furthermore, docetaxel has been shown to bind to human plasma proteins (see, for example, Urien et al., Invest. New Drugs, 14(b), 147-51 (1996)).

[0146] mTOR inhibitors (e.g., limus-based drugs, e.g., rapamycin or its derivatives) are "stabilized" in aqueous suspensions for extended periods, e.g., at least about 0.1, 0.2, 0.25, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 24, 36, 48, 60, or 72 hours, if left suspended in an aqueous medium (e.g., without visible precipitation or sedimentation). The suspension is generally suitable for administration to an individual (e.g., a human), but not necessarily. The stability of the suspension is generally assessed (but not essential) at the storage temperature (e.g., room temperature (e.g., 20-25°C) or refrigerated conditions (e.g., 4°C)). For example, a suspension is stable at the storage temperature if, after observation for about 15 minutes after preparation, it does not show visible aggregation or particle aggregation to the naked eye, or if observed 1000 times using a light microscope. Stability can also be evaluated under accelerated testing conditions, such as temperatures above approximately 40°C.

[0147] The compositions described herein may be stable aqueous suspensions of mTOR inhibitors at any of the following concentrations: about 0.1 to about 200 mg / ml, about 0.1 to about 150 mg / ml, about 0.1 to about 100 mg / ml, about 0.1 to about 50 mg / ml, about 0.1 to about 20 mg / ml, about 1 to about 10 mg / ml, about 2 mg / ml to about 8 mg / ml, about 4 to about 6 mg / ml, and about 5 mg / ml. In some embodiments, the concentration of the mTOR inhibitor is at least one of the following: 0.2 mg / ml, 1.3 mg / ml, 1.5 mg / ml, 2 mg / ml, 3 mg / ml, 4 mg / ml, 5 mg / ml, 6 mg / ml, 7 mg / ml, 8 mg / ml, 9 mg / ml, 10 mg / ml, 15 mg / ml, 20 mg / ml, 25 mg / ml, 30 mg / ml, 40 mg / ml, 50 mg / ml, 100 mg / ml, 150 mg / ml, or 200 mg / ml.

[0148] In some embodiments, albumin is present in an amount sufficient to stabilize an mTOR inhibitor (e.g., a limus-based drug, e.g., rapamycin or its derivatives) in an aqueous suspension of a certain concentration. For example, the concentration of the mTOR inhibitor (e.g., a limus-based drug, e.g., rapamycin or its derivatives) in the composition is about 0.1 to about 100 mg / ml, including, for example, any of the following: about 0.1 to about 50 mg / ml, about 0.1 to about 20 mg / ml, about 1 to about 10 mg / ml, about 2 mg / ml to about 8 mg / ml, about 4 to about 6 mg / ml, or about 5 mg / ml. In some embodiments, the concentration of the mTOR inhibitor (e.g., a limus-based drug, e.g., rapamycin or its derivatives) is at least one of the following: 1.3 mg / ml, 1.5 mg / ml, 2 mg / ml, 3 mg / ml, 4 mg / ml, 5 mg / ml, 6 mg / ml, 7 mg / ml, 8 mg / ml, 9 mg / ml, 10 mg / ml, 15 mg / ml, 20 mg / ml, 25 mg / ml, 30 mg / ml, 40 mg / ml, and 50 mg / ml. In some embodiments, albumin is present in an amount that avoids the use of surfactants (e.g., cremophor) so that the composition is free of or substantially free of surfactants (e.g., cremophor).

[0149] In some embodiments, the composition in liquid form contains about 0.1% to about 50% (w / v) of albumin (e.g., about 0.5% (w / v), about 5% (w / v), about 10% (w / v), about 15% (w / v), about 20% (w / v), about 30% (w / v), about 40% (w / v), or about 50% (w / v)).

[0150] In some embodiments, albumin allows the composition to be administered to an individual (e.g., a human) without significant side effects. In some embodiments, albumin (e.g., human serum albumin or human albumin) is in an effective amount to reduce one or more side effects of administering an mTOR inhibitor (e.g., a limus-based drug, e.g., rapamycin or its derivatives) to a human. The term "reduce one or more side effects" of administering an mTOR inhibitor (e.g., a limus-based drug, e.g., rapamycin or its derivatives) refers to the reduction, mitigation, elimination or avoidance of one or more undesirable effects by the mTOR inhibitor, as well as side effects by the delivery vehicle used to deliver the mTOR inhibitor (e.g., a solvent that makes the limus-based drug suitable for injection). Such side effects include, for example, myelosuppression, neurotoxicity, hypersensitivity, inflammation, venous irritation, phlebitis, pain, skin irritation, peripheral neuropathy, neutropenic fever, anaphylactic reaction, venous thrombosis, extravasation, and combinations thereof. However, these side effects are merely illustrative, and other side effects or combinations of side effects associated with limousine drugs (e.g., limousine drugs such as rapamycin or its derivatives) can be reduced.

[0151] In some embodiments, the composition is a dry (e.g., lyophilized) composition that can be reconstituted, resuspended, or rehydrated to generally form a stable aqueous suspension of nanoparticles containing an mTOR inhibitor and albumin. In some embodiments, the composition is a liquid (e.g., aqueous) composition obtained by reconstituting or resuspending the dry composition. In some embodiments, the composition is an intermediate liquid (e.g., aqueous) composition that can be dried (e.g., lyophilized).

[0152] A. mTOR inhibitors In some embodiments, the methods described herein involve the administration of a nanoparticle composition of an mTOR inhibitor. As used herein, “mTOR inhibitor” refers to an inhibitor of mTOR. mTOR is a serine / threonine-specific protein kinase downstream of the phosphatidylinositol 3-kinase (PI3K) / Akt (protein kinase B) pathway and is a major regulator of cell survival, proliferation, stress, and metabolism. It is found in many human cancers, and mTOR inhibition has resulted in substantial inhibitory effects on tumor progression.

[0153] Mammalian rapamycin targets (mTORs) (also known as the mechanistic targets of rapamycin or FK506-binding protein 12-rapamycin-associated protein 1 (FRAP1)) are atypical serine / threonine protein kinases present in two distinct complexes, mTOR complex 1 (mTORC1) and mTOR complex 2 (mTORC2). mTORC1 is composed of mTOR, mTOR regulatory-associated protein (Raptor), lethal mammalian protein SEC13 protein 8 (MLST8), PRAS40, and DEPTOR (Kim et al. (2002). Cell 110:163-75, Fang et al. (2001). Science 294(5548):1942-5). mTORC1 incorporates four major signaling inputs: nutrients (e.g., amino acids and phosphatidic acid), growth factors (insulin), energy, and stress (e.g., hypoxia and DNA damage). Amino acid availability is signaled to mTORC1 via a pathway involving lag-and-lagtor (LAMTOR1-3) growth factors, and hormones (e.g., insulin) are signaled to mTORC1 via Akt, which inactivates TSC2 and prevents inhibition of mTORC1. Alternatively, low ATP levels trigger AMPK-dependent activation of TSC2 and phosphorylation of lapta to reduce mTORC1 signaling proteins.

[0154] Active mTORC1 has multiple downstream biological effects, including mRNA translation via phosphorylation of downstream targets (4E-BP1 and p70 S6 kinase), repression of autophagy (Atg13, ULK1), ribosome biosynthesis, and transcriptional activation leading to mitochondrial metabolism or adipogenesis. Therefore, mTORC1 activity promotes cell proliferation under favorable conditions or accelerates catabolic processes during stress under unfavorable conditions.

[0155] mTORC2 is composed of mTOR, mTOR's rapamycin-insensitive companion (RICTOR), GβL, and mammalian stress-activated protein kinase interacting protein 1 (mSIN1). In contrast to mTORC1, many upstream signaling and cellular functions are defined (see above), relatively little is known about the biology of mTORC2. mTORC2 regulates the cytoskeleton through its stimulation of F-actin stress filaments, paxilin, RhoA, Rac1, Cdc42, and protein kinase Cα (PKCα). Knockdown of mTORC2 components has been observed to affect actin polymerization and disrupt cell morphology (Jacinto et al. (2004). Nat. Cell Biol. 6, 1122-1128, Sarbassov et al. (2004). Curr. Biol. 14, 1296-1302). This suggests that mTORC2 regulates the actin cytoskeleton by promoting protein kinase Cα (PKCα) phosphorylation, paxilin phosphorylation and its relocalization to local adhesion, and GTP loading of RhoA and Rac1. The molecular mechanisms by which mTORC2 modulates these processes remain unclear.

[0156] In some embodiments, the mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or its derivatives) is an mTORC1 inhibitor. In some embodiments, the mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or its derivatives) is an mTORC2 inhibitor. In some embodiments, the mTOR inhibitor (e.g., a limus-based drug, e.g., sirolimus or its derivatives) is an inhibitor of both mTORC1 and mTORC2.

[0157] In some embodiments, the mTOR inhibitor is a limousine drug, including sirolimus and its analogs. Examples of limousine drugs include, but are not limited to, temsirolimus (CCI-779), everolimus (RAD001), lidafololimus (AP-23573), defololimus (MK-8669), zotarolimus (ABT-578), pimecrolimus, and tacrolimus (FK-506). In some embodiments, the limousine drug is selected from the group consisting of temsirolimus (CCI-779), everolimus (RAD001), lidafololimus (AP-23573), defololimus (MK-8669), zotarolimus (ABT-578), pimecrolimus, and tacrolimus (FK-506). In some embodiments, the mTOR inhibitor is an mTOR kinase inhibitor, such as CC-115 or CC-223.

[0158] In some embodiments, the mTOR inhibitor is sirolimus. Sirolimus is a macrolide antibiotic that inhibits the mTOR pathway by conjugating with FKBP-12 and binding to mTORC1.

[0159] In some embodiments, the mTOR inhibitor is sirolimus (rapamycin), BEZ235 (NVP-BEZ235), everolimus (also known as RAD001, Zortress, Certican, and Afinitor), AZD8055, temsirolimus (also known as CCI-779 and Torisel), CC-115, CC-223, PI-103, Ku-0063794, INK 128, AZD2014, NVP-BGT226, PF-04691502, CH5132799, GDC-0980 (RG7422), Torin 1. Selected from the group consisting of YAW-600, WYE-125132, WYE-687, GSK2126458, PF-05212384 (PKI-587), PP-121, OSI-027, Palomid 529, PP242, XL765, GSK1059615, WYE-354, and ridafololimus (also known as defololimus).

[0160] BEZ235 (NVP-BEZ235) is an imidazoquinoline derivative that acts as an mTORC1 catalytic inhibitor (Roper J, et al. PLoS One, 2011, 6(9), e25132). Everolimus is a 40-O-(2-hydroxyethyl) derivative of sirolimus and binds to cyclophylline FKBP-12, and this complex also binds to mTORC1. AZD8055 is a small molecule that inhibits the phosphorylation of mTORC1 (p70S6K and 4E-BP1). Temsirolimus is a small molecule that forms a complex with FK506-binding protein and, if it remains in the mTORC1 complex, prevents mTOR activation. PI-103 is a small molecule that inhibits the activation of the rapamycin-sensitive (mTORC1) complex (Knight et al. (2006) Cell. 125:733-47). KU-0063794 is a small molecule that dose- and time-dependently inhibits the phosphorylation of mTORC1 at Ser2448. INK 128, AZD2014, NVP-BGT226, CH5132799, and WYE-687 are small molecule inhibitors of mTORC1. PF-04691502 inhibits mTORC1 activity. GDC-0980 is an orally bioavailable small molecule that inhibits class I PI3 kinase and TORC1. Torin 1 is a potent small molecule inhibitor of mTOR. WAY-600 is a potent ATP competitive and selective inhibitor of mTOR. WYE-125132 is an ATP competitive small molecule inhibitor of mTORC1. GSK2126458 is an inhibitor of mTORC1. PKI-587 is a very potent dual inhibitor of PI3Kα, PI3Kγ, and mTOR. PP-121 is a multi-target inhibitor of PDGF, Hck, mTOR, VEGFR2, Src, and Abl. OSI-027 is a selective and potent dual inhibitor of mTORC1 and mTORC2, with IC50s of 22nM and 65nM, respectively. Palomid 529 is a small molecule inhibitor of mTORC1 that lacks affinity for ABCB1 / ABCG2 and has good brain permeability (Lin et al. (2013) Int J Cancer DOI:10.1002 / ijc.28126 (electronic publication before print)). PP242 is a selective mTOR inhibitor.XL765 is a dual mTOR / PI3k inhibitor against mTOR, p110α, p110β, p110γ, and p110δ. GSK1059615 is a novel dual inhibitor of PI3Kα, PI3Kβ, PI3Kδ, PI3Kγ, and mTOR. WYE-354 inhibits mTORC1 in HEK293 cells (0.2 μM~5 μM) and HUVEC cells (10 nM~1 μM). WYE-354 is a potent and specific ATP-competitive inhibitor of mTOR. Defololimus (Ridaforolimus, AP23573, MK-8669) is a selective mTOR inhibitor.

[0161] C. Other components in the nanoparticle composition In some embodiments, the composition is suitable for administration to humans. In some embodiments, the composition is suitable for administration to mammals, such as domestic pets and farm animals, for example, in a veterinary context. The following formulations and methods are merely illustrative and not limiting. Formulations suitable for oral administration may consist of (a) a liquid solution such as an effective amount of the compound dissolved in a diluent such as water, saline, or orange juice; (b) capsules, sachets, or tablets containing a predetermined amount of the active ingredient as a solid or granular preparation, respectively; (c) a suspension in a suitable liquid; and (d) a suitable emulsion. Tablet forms may contain lactose, mannitol, corn starch, potato starch, microcrystalline cellulose, gum arabic, gelatin, colloidal silicon dioxide, croscarmellose sodium, talc, magnesium stearate, stearic acid, and one or more other excipients, colorants, diluents, buffers, wetting agents, preservatives, flavorings, and pharmacologically suitable excipients. Lozenge forms can contain the active ingredient in flavorings, usually sucrose and acacia or tragacanth, as well as lozenges containing the active ingredient in an inert base such as gelatin and glycerin, or sucrose and acacia, emulsions, gels, etc., containing the active ingredient plus excipients, which are known in the art.

[0162] Suitable carriers, excipients, and diluents include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, saline solution, syrup, methylcellulose, methyl hydroxybenzoate and propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil. The formulation may further contain lubricants, wetting agents, emulsifiers and suspending agents, preservatives, sweeteners, or flavoring agents.

[0163] Suitable formulations for parenteral administration include aqueous and non-aqueous isotonic sterile injection solutions that may contain antioxidants, buffers, bacteriostatic agents, and solutes to ensure compatibility with the recipient's blood, as well as aqueous and non-aqueous sterile suspensions that may contain suspending agents, solubilizers, thickeners, stabilizers, and preservatives. The formulations can be provided in sealed containers of unit or multiple doses, such as ampoules and vials, and can be stored in a freeze-dried state requiring only the addition of a sterile liquid excipient, such as water, immediately before use for injection. Immediate injection solutions and suspensions can be prepared from the aforementioned types of sterile powders, granules, and tablets. Injectable formulations are preferred.

[0164] In some embodiments, the composition is formulated to have a pH range of about 4.5 to about 9.0, including, for example, one of the pH ranges of about 5.0 to about 8.0, about 6.5 to about 7.5, and about 6.5 to about 7.0. In some embodiments, the pH of the composition is formulated to be about 6 or higher, including, for example, one or more of about 6.5, 7, or 8 (e.g., about 8). The composition can also be made isotonic with blood by the addition of a suitable tonicity modifier such as glycerol.

[0165] D. Rapamycin albumin-based nanoparticle composition The methods described herein are particularly suitable for albumin-based nanoparticle compositions as described in more detail herein. In some embodiments, the nanoparticle compositions include (a) nanoparticles containing rapamycin and albumin, and (b) non-nanoparticle portions containing rapamycin and albumin. The rapamycin and albumin in the nanoparticles associate with each other within the nanoparticles. For example, the nanoparticles may include a coating having albumin surrounding a core containing rapamycin. In the non-nanoparticle portions of the composition, rapamycin and albumin may or may not associate with each other (i.e., rapamycin may be in a reversible binding equilibrium with albumin), but they do not associate with each other to form nanoparticles. That is, the nanoparticle composition may include nanoparticle-bound albumin and nanoparticle-bound rapamycin in the nanoparticle portions of the composition, and non-nanoparticle albumin and non-nanoparticle rapamycin in the non-nanoparticle portions of the composition. As used herein, “in nanoparticles” is used synonymously with “in nanoparticle portions.” Albumin in nanoparticles may be further distinguishable from albumin in the non-nanoparticle portion of the composition; for example, the oligomeric profile of albumin in nanoparticles may differ from the oligomeric profile of albumin in the non-nanoparticle portion of the composition. The oligomeric profile refers to the percentage of various albumin species compared to the total albumin in the composition. The types of albumin species include albumin monomers, dimers, trimers, oligomers, and polymers.As used herein, “albumin monomer” or “monomeralbumin” refers to albumin species having one and only one albumin unit; “albumin dimer” or “dimeralbumin” refers to albumin species having two and only two albumin units; “albumin trimer” or “trimeralbumin” refers to albumin species having three and only three albumin units; “high molecular weight albumin” refers to albumin species having a higher molecular weight than albumin monomers and albumin dimers; and “albumin oligomer” or “oligomeric albumin” refers to low molecular weight high molecular weight albumin species associated with a UV-based size exclusion chromatography peak observed between the peak associated with albumin dimers and the high molecular weight high molecular weight high molecular weight albumin species.

[0166] The albumin nanoparticles associate with the rapamycin nanoparticles so that the nanoparticle suspension has a high concentration of rapamycin, thereby enabling the composition to be used as a pharmaceutical composition for treating certain diseases, such as cancer. The manufactured nanoparticles (for example, those that can be produced using the method described herein) can be formulated, filtered, or otherwise processed to obtain a pharmaceutical composition suitable for medical use in human organisms.

[0167] Generally, to produce the rapamycin pharmaceutical compositions described herein, rapamycin is dissolved in an organic solvent. Suitable organic solvents include, for example, ketones, esters, ethers, chlorinated solvents, and other solvents known in the art. For example, the organic solvent may be a mixture of methylene chloride / ethanol, chloroform / ethanol, or chloroform / tert-butanol (e.g., any one of the ratios about 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1 or 9:1, or any one of the ratios 3:7, 5:7, 4:6, 5:5, 6:5, 8:5, 9:5, 9.5:5, 5:3, 7:3, 6:4 or 9.5:0.5). In some embodiments, the organic solvent contains about 10% to about 50% by volume of tert-butanol. In some embodiments, the organic solvent contains tert-butanol in any of the following amounts: about 10% by volume, 15% by volume, 20% by volume, 25% by volume, 30% by volume, 35% by volume, 40% by volume, 45% by volume, or 50% by volume. In some embodiments, the organic solvent contains tert-butanol in any of the following amounts: about 10–15% by volume, 15–20% by volume, 20–25% by volume, 25–30% by volume, 30–35% by volume, 35–40% by volume, 40–45% by volume, or 45–50% by volume, or any combination of such ranges. In some embodiments, the organic solvent contains chloroform in amounts of about 50% by volume to about 90% by volume. In some embodiments, the organic solvent contains chloroform in any of the following amounts: about 50% by volume, 55% by volume, 60% by volume, 65% by volume, 70% by volume, 75% by volume, 80% by volume, 85% by volume, or 90% by volume. In some embodiments, the organic solvent comprises chloroform in an amount of about 50–55% by volume, 55–60% by volume, 60–65% by volume, 65–70% by volume, 70–75% by volume, 75–80% by volume, 80–85% by volume, or 85–90% by volume, or any combination of such ranges. In some embodiments, the organic solvent comprises about 10% to about 50% by volume of tert-butanol and about 50% to about 90% by volume of chloroform.In some embodiments, the organic solvent comprises chloroform and tert-butanol in a volume ratio of about 1:1 to about 1:9, for example, about 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, and 9:1.

[0168] Albumin (for example, recombinant albumin, for example, NOVOZYME® recombinant albumin or INTRIVIA as disclosed herein) (商標)Recombinant albumin is dissolved in an aqueous solution (e.g., water) and combined with rapamycin solution to form a crude emulsion. The mixture is subjected to high-pressure homogenization (e.g., Avestin, APV Gaulin, MICROFLUIDIZER®, e.g., Microfluidics, Stansted or Ultra Turrax homogenizers, or MICROFLUIDIZER® processor M-110EH). The emulsion may be circulated through a high-pressure homogenizer for about 2 to about 100 cycles, e.g., about 5 to about 50 cycles or about 6 to about 20 cycles (e.g., any one of about 6, 8, 10, 12, 14, 16, 18 or 20 cycles). The organic solvent can then be removed by evaporation using known suitable apparatus for removal purposes (including, but not limited to, rotary evaporators, drip-film evaporators, wiped-film evaporators, spray dryers, etc., which can be operated in batch mode or continuous operation). In some embodiments, the evaporator is a wiped-film evaporator. The solvent can be removed under reduced pressure (e.g., any one of approximately 25 mmHg, 30 mmHg, 40 mmHg, 50 mmHg, 100 mmHg, 200 mmHg, or 300 mmHg). The duration used to remove the solvent under reduced pressure can be adjusted based on the volume of the formulation. For example, for a formulation produced on a 300 mL scale, the solvent can be removed under approximately 1 to approximately 300 mmHg (e.g., any one of approximately 5 to 100 mmHg, 10 to 50 mmHg, 20 to 40 mmHg, or 25 mmHg) for approximately 5 to approximately 60 minutes (e.g., any one of approximately 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 18, 20, 25, or 30 minutes). The resulting dispersion can be further lyophilized.

[0169] The nanoparticle compositions described herein (e.g., pharmaceutical compositions) include (1) an oligomeric state of albumin associated with nanoparticles (e.g., in nanoparticles), for example, a percentage of albumin monomers, dimers, and / or polymers (or trimers) of albumin associated with nanoparticles (e.g., in nanoparticles), and (2) an oligomeric state of albumin associated with the non-nanoparticle portion of the composition (e.g., in the non-nanoparticle portion), for example, albumin monomers, dimers, and / or polymers (and (1) Percentage of albumin (trimer), (2) Oligomeral state of total albumin in the composition, e.g., percentage of albumin monomers, dimers and / or polymers (or trimers) of total albumin in the composition, (3) Particle size profile of nanoparticles, e.g., average particle size, polydispersity index and / or particle size distribution, (4) Percentage of nanoparticles that are albumin (e.g., weight percentage) and / or percentage of nanoparticles that are rapamycin (e.g., weight percentage), (5) Weight ratio of albumin to rapamycin in nanoparticles, (6) Percentage of rapamycin in the non-nanoparticle portion of the composition (8) the weight ratio of albumin to rapamycin in the non-nanoparticle portion of the composition, (9) the weight ratio of total albumin to total rapamycin in the composition, (10) the portion of rapamycin in the nanoparticles (or non-nanoparticle portion of the composition) compared to total rapamycin in the composition (e.g., weight percent), (11) the portion of albumin in the non-nanoparticle portion (or nanoparticles) compared to total albumin in the composition (e.g., weight percent), (12) the concentration of albumin in the composition, (13) the amount of albumin in the non-nanoparticle portion of the composition (14) Concentration of albumin in the composition associated with nanoparticles (e.g., in nanoparticles), (15) Concentration of rapamycin in the composition, (16) Concentration of rapamycin in the non-nanoparticle portion of the composition, (17) Concentration of rapamycin in the composition associated with nanoparticles (e.g., in nanoparticles), (18) Osmolality of the composition, (19) Viscosity of the composition, (20) pH of the composition, (21) Stability of nanoparticles in the composition, (22) Amount of residual solvent in the composition, (23) Zeta potential of nanoparticles in the composition, (24) Crystalline state of rapamycin in nanoparticles, (25) Shape,The nanoparticles may have distinct properties with respect to one or more of the following (in any combination): (26) the particle morphology of the nanoparticles, such as sphericity and coating thickness, and / or surface area to volume ratio; (27) the weight percentage of seco-rapamycin in the nanoparticles, on a weight basis, compared to the total weight of seco-rapamycin and rapamycin; (28) the presence, percentage, or concentration of albumin stabilizers (e.g., sodium caprylate and / or N-acetyltryptophanate) in the composition; (29) the recovery of rapamycin after filtration; (30) the in vitro release kinetics of the nanoparticles; and (31) the portion of total rapamycin in the composition that is in the non-nanoparticle portion of the composition and not bound to albumin, and / or (31) the weight percentage of seco-rapamycin in the composition, on a weight basis, compared to the total weight of seco-rapamycin and rapamycin. In some embodiments, the oligomeric state of nanoparticles, non-nanoparticle portions, or the entire composition (e.g., percentages of albumin monomers, dimers, or polymers (or trimers)) is evaluated by size exclusion chromatography using a saline mobile phase coupled to a multi-angle light scattering (MALS) detector.

[0170] The nanoparticle compositions described herein (such pharmaceutical compositions) include: (1) a percentage of albumin oligomers (e.g., in the nanoparticles) associated with nanoparticles, e.g., albumin monomers, dimers, oligomers, and / or polymers (other than oligomers) of albumin associated with nanoparticles (e.g., in the nanoparticles); and (2) an oligomer of albumin associated with the non-nanoparticle portion of the composition, e.g., albumin monomers, dimers, oligomers, and / or polymers (other than oligomers) of albumin associated with the non-nanoparticle portion of the composition. (3) Percentage of albumin monomers, dimers, oligomers, and / or polymers (other than oligomers) of total albumin in the composition, e.g., percentage of albumin monomers, dimers, oligomers, and / or polymers (other than oligomers) of total albumin in the composition, (4) Particle size profile of nanoparticles, e.g., average particle size, polydispersity index, and / or particle size distribution, (5) Percentage of nanoparticles that are albumin (e.g., by weight), and / or Percentage of nanoparticles that are rapamycin (e.g., by weight), (6) Percentage of albumin relative to rapamycin in nanoparticles (7) Weight ratio of albumin to rapamycin in the non-nanoparticle portion of the composition, (8) Weight ratio of albumin to rapamycin in the non-nanoparticle portion of the composition, (9) Weight ratio of total albumin to total rapamycin in the composition, (10) Percentage of rapamycin in nanoparticles (or the non-nanoparticle portion of the composition) compared to total rapamycin in the composition (e.g., weight percentage), (11) Percentage of albumin in the non-nanoparticle portion (or nanoparticles) compared to total albumin in the composition (e.g., weight percentage), (12) A (13) Concentration of albumin in the non-nanoparticle portion of the composition, (14) Concentration of albumin in the composition associated with nanoparticles (e.g., in the nanoparticles), (15) Concentration of rapamycin in the composition, (16) Concentration of rapamycin in the non-nanoparticle portion of the composition, (17) Concentration of rapamycin in the composition associated with nanoparticles (e.g., in the nanoparticles), (18) Osmolality of the composition, (19) Viscosity of the composition, (20) pH of the composition, (21) Stability of nanoparticles in the composition, (22) Amount of residual solvent in the composition, (23) Zeta potential of nanoparticles in the composition.The composition may have distinct properties with respect to one or more of the following (in any combination): (24) the crystalline state of rapamycin in the nanoparticles; (25) the particle morphology of the nanoparticles, such as shape, sphericity, and coating thickness, and / or surface area to volume ratio; (26) the weight percentage of seco-rapamycin in the nanoparticles, on a weight basis, compared to the sum of seco-rapamycin and rapamycin; (27) the presence, percentage, or concentration of albumin stabilizers (e.g., sodium caprylate and / or N-acetyltryptophanate) in the composition; (28) the recovery of rapamycin after filtration; (29) the in vitro release kinetics of the nanoparticles; (30) the portion of total rapamycin in the composition that is in the non-nanoparticle portion of the composition and not bound to albumin, and / or (31) the weight percentage of seco-rapamycin in the composition, on a weight basis, compared to the sum of seco-rapamycin and rapamycin. As used herein, “albumin oligomer” or “oligomeric albumin” refers to a lower molecular weight polymer albumin species associated with a UV absorption-based size exclusion chromatography peak observed between the peak associated with the albumin dimer and the higher molecular weight polymer albumin species. In some embodiments, the oligomeric state of the nanoparticles, non-nanoparticle portions, or the entire composition (e.g., percentages of albumin monomers, dimers, oligomers, or polymers (non-oligomeric)) is evaluated by size exclusion chromatography using a mobile phase (e.g., aqueous buffer containing 7.5% methanol) containing the aqueous portion and the miscible organic portion, coupled to a UV detector. In some embodiments, the percentage of albumin in the nanoparticle portions in the form of monomers, dimers, oligomers, or polymer albumins (non-oligomeric albumins) is determined by separating the nanoparticles from the non-nanoparticle portion, dissolving the nanoparticles, and subjecting the dissolved nanoparticles to size exclusion chromatography. In some embodiments, size exclusion chromatography uses a mobile phase containing an aqueous moiety and a miscible organic moiety (e.g., an aqueous buffer containing 7.5% methanol) in combination with a UV detector.

[0171] In some embodiments, the nanoparticle composition is such that (1) about 80% to about 95% of the total albumin in the composition (or as further provided herein) is in the form of monomeric albumin; (2) about 4% to about 15% of the total albumin in the composition (or as further provided herein) is in the form of dimeric albumin; (3) about 0.5% to about 5% of the total albumin in the composition (or as further provided herein) is in the form of polymeric albumin (or trimeric albumin); (4) the weight ratio of total albumin to total rapamycin in the composition is about 1:1 to about 10:1 (or as further provided herein); (5) about 90% or more of the total rapamycin in the composition (or as further provided herein) is in nanoparticles; and (6) about 90% or more of the total albumin in the composition (or as further provided herein) is non-nanoparticle nanoparticles. The composition is located in the sub-portion and has one or more distinct properties: (7) the composition comprises tert-butanol at a concentration of less than about 10 μg / mL or less than about 10 ppm (or as further provided herein); (8) the composition comprises chloroform at a concentration of less than about 5 μg / mL or less than about 5 ppm (or as further provided herein); (9) the composition comprises an albumin stabilizer (e.g., sodium caprylate and / or N-acetyltryptophanate); (10) at least about 80% or more of the rapamycin in the composition (or as further provided herein) is recoverable after filtering the composition through a 0.2 micron filter; (11) the composition is stable for at least 24 hours; and / or (12) less than about 5% of the total rapamycin in the composition is located in the non-nanoparticle portion of the composition and does not bind to albumin in the non-nanoparticle portion of the composition.In some embodiments, the nanoparticle composition may be a nanoparticle suspension, and the nanoparticle composition may have one or more distinct properties (in addition to or instead of any one of the aforementioned distinct properties) such as: (1) a concentration of albumin in the composition of about 30 mg / mL to about 100 mg / mL (or as further provided herein); (2) a concentration of rapamycin in the composition of about 1 mg / mL to about 15 mg / mL (or as further provided herein, for example, about 1 mg / mL to about 7 mg / mL); (3) an osmolality of the composition of about 300 mOsm / kg to about 350 mOsm / kg (or as separately provided herein); (4) a viscosity of the composition of about 1.2 cP to about 1.5 cP (or as separately provided herein); and / or (5) a pH of the composition of about 6.0 to about 7.5 (or as separately provided herein).

[0172] In some embodiments, the nanoparticles of the composition are: (1) about 70% to about 85% of the albumin in the nanoparticles (or as otherwise provided herein) in the form of albumin monomers; (2) about 9% to about 20% of the albumin in the nanoparticles (or as otherwise provided herein) in the form of albumin dimers; (3) about 5% to about 15% of the albumin in the nanoparticles (or as otherwise provided herein) in the form of albumin polymers (or albumin trimers); and (4) the nanoparticles are about 200 nm or less (or as otherwise provided herein). (5) The nanoparticles have a volume-weighted average particle size and / or Z-average particle size of approximately 50 nm to approximately 200 nm, (6) the nanoparticles have a polydispersity index of less than approximately 0.2 (or approximately 0.03 to approximately 0.2, as provided separately herein), (7) the nanoparticles are approximately 25% to approximately 45% by weight of albumin (or as provided separately herein), and (8) the nanoparticles are approximately 55% to approximately 75% by weight of albumin. (9) the weight ratio of albumin to rapamycin in the nanoparticles is about 1:1 to about 1:4 (or as otherwise provided herein), (10) the zeta potential of the nanoparticles in the composition is about -25mV to about -50mV (or as otherwise provided herein), (11) the nanoparticles are amorphous, (12) the rapamycin in the nanoparticles is amorphous, (13) the vinyl chain of rapamycin in the nanoparticles interacts with the albumin in the nanoparticles, (14) the composition is (15) The nanoparticles contain less than 2.5% seco-rapamycin by weight (or, as provided separately herein, for example, about 0.2% to about 2.5%) compared to the sum of seco-rapamycin and rapamycin, and / or (16) The composition contains less than 3% by weight of seco-rapamycin (or, as provided separately herein, for example, about 0.2% to about 2%) compared to the sum of seco-rapamycin and rapamycin.It has one or more distinct properties, including containing 5%. In some embodiments, the nanoparticle composition may be a nanoparticle suspension, and in some embodiments, the concentration of albumin in the nanoparticle suspension contained in the nanoparticles is about 1.8 mg / mL to about 3 mg / mL (or as otherwise provided herein).

[0173] In some embodiments, the nanoparticles of the composition consist of (1) about 25% to about 50% of the albumin in the nanoparticles (or as otherwise provided herein) in the form of albumin monomers, (2) about 5% to about 16% of the albumin in the nanoparticles (or as otherwise provided herein) in the form of albumin dimers, (3) about 1% to about 4.5% of the albumin in the nanoparticles (or as otherwise provided herein) in the form of albumin polymers, and (4) about 42% to about 60% of the albumin in the nanoparticles (or as otherwise provided herein). (As separately provided in the Specification) the albumin polymer (other than oligomers) is in the form of (5) nanoparticles having a volume-weighted average particle size and / or Z-average particle size of about 200 nm or less (or, as separately provided herein, for example, about 50 nm to about 200 nm), (6) nanoparticles having a polydispersity index of less than about 0.2 (or, as separately provided herein, for example, about 0.03 to about 0.2), and (7) the span of the particle size distribution ((Dv95-Dv5) / Dv50) is about 0.8 to about 1.2 (or, as separately provided herein) (8) The nanoparticles are about 25% to about 45% by weight of albumin (or as otherwise provided herein), (9) The nanoparticles are about 55% to about 75% by weight of rapamycin (or as otherwise provided herein), (10) The weight ratio of albumin to rapamycin in the nanoparticles is about 1:1 to about 1:4 (or as otherwise provided herein), (11) The zeta potential of the nanoparticles in the composition is about -25mV to about -50mV (or as otherwise provided herein), (12 (13) The nanoparticles have an amorphous form, (14) The vinyl chain of rapamycin in the nanoparticles interacts with the albumin in the nanoparticles, (15) At least a portion of the nanoparticles in the composition (e.g., at least 20%, or as otherwise provided herein) are non-spherical, and (16) The nanoparticles contain less than about 2.5% seco-rapamycin by weight (or, as otherwise provided herein, for example, about 0.2% to about 2%) compared to the sum of seco-rapamycin and rapamycin.(17) The composition contains 5%, and / or (17) the composition has one or more distinct properties, including containing less than 3% by weight of seco-rapamycin (or, as otherwise provided herein, for example, about 0.2% to about 3%) compared to the sum of seco-rapamycin and rapamycin. In some embodiments, the nanoparticle composition may be a nanoparticle suspension, and in some embodiments, the concentration of albumin in the nanoparticle suspension contained in the nanoparticles is about 1.8 mg / mL to about 3 mg / mL (or as otherwise provided herein).

[0174] In some embodiments, the non-nanoparticle portion of the composition has one or more distinct properties: (1) about 80% to about 95% (or as otherwise provided herein) of the albumin in the non-nanoparticle portion of the composition is in the form of albumin monomers; (2) about 5% to about 14% (or as otherwise provided herein) of the albumin in the non-nanoparticle portion of the composition is in the form of albumin dimers; and / or (3) about 1% to about 5% (or as otherwise provided herein) of the albumin in the non-nanoparticle portion of the composition is in the form of albumin polymers (or albumin trimers). In some embodiments, the nanoparticle composition may be a nanoparticle suspension, and the non-nanoparticle portion of the nanoparticle suspension may have one or more distinct properties (in addition to or instead of any one of the distinct properties described above) such that (1) the concentration of albumin in the non-nanoparticle portion of the composition is about 30 mg / mL to about 100 mg / mL (or as separately provided herein), and / or (2) the concentration of rapamycin in the non-nanoparticle portion is about 20 μg / mL to about 55 μg / mL (or as separately provided herein).

[0175] In some embodiments, the non-nanoparticle portion of the composition has one or more of the following characteristics: (1) about 80% to about 95% of the albumin in the non-nanoparticle portion of the composition (or as otherwise provided herein) is in the form of albumin monomers; (2) about 5% to about 16% of the albumin in the non-nanoparticle portion of the composition (or as otherwise provided herein) is in the form of albumin dimers; about 0.5% to about 4% of the albumin in the non-nanoparticle portion of the composition (or as otherwise provided herein) is in the form of albumin oligomers; and / or (4) about 0.5% to about 3% of the albumin in the non-nanoparticle portion of the composition (or as otherwise provided herein) is in the form of albumin polymers (other than oligomers). In some embodiments, the nanoparticle composition may be a nanoparticle suspension, and the non-nanoparticle portion of the nanoparticle suspension may have one or more distinct properties (in addition to or instead of any one of the distinct properties described above) such that (1) the concentration of albumin in the non-nanoparticle portion of the composition is about 30 mg / mL to about 100 mg / mL (or as separately provided herein), and / or (2) the concentration of rapamycin in the non-nanoparticle portion is about 20 μg / mL to about 55 μg / mL (or as separately provided herein).

[0176] The compositions described herein (e.g., pharmaceutical compositions) may be in liquid form (e.g., as a nanoparticle suspension) or powder form. For example, in some embodiments, the composition is a liquid nanoparticle suspension (e.g., before lyophilization). In some embodiments, the composition is a reconstituted suspension (e.g., in an aqueous solution such as physiological saline). In some embodiments, the composition is dried, for example, by lyophilization. In some embodiments, the composition is sterile. In some embodiments, the composition is contained in a sealed container such as a sealed vial (e.g., a glass vial) or a sealed bag.

[0177] In some embodiments, the nanoparticle composition comprises (a) nanoparticles containing rapamycin and albumin (e.g., human albumin), and (b) a non-nanoparticle portion containing albumin (e.g., human albumin) and rapamycin. In some embodiments, about 0.5% to about 5% of the albumin in the non-nanoparticle portion or the total albumin in the nanoparticle composition is in the form of high molecular weight albumin (or trimer albumin). In some embodiments, about 4% to about 14% of the albumin in the non-nanoparticle portion or the total albumin in the nanoparticle composition is in the form of dimer albumin. In some embodiments, about 80% to about 95% of the albumin in the non-nanoparticle portion or the total albumin in the nanoparticle composition is in the form of monomer albumin. In some embodiments, the weight ratio of albumin to rapamycin in the composition is about 1:1 to about 10:1. In some embodiments, about 90% or more of the albumin in the composition is in the non-nanoparticle portion. In some embodiments, about 90% or more of the rapamycin in the composition is in nanoparticles. In some embodiments, the concentration of albumin in the nanoparticle composition in the non-nanoparticle portion or the total albumin concentration in the nanoparticle composition is about 30 mg / mL to about 100 mg / mL. In some embodiments, the osmolality of the composition is about 300 mOsm / kg to about 350 mOsm / kg. In some embodiments, the viscosity of the composition is about 1.2 cP to about 1.5 cP. In some embodiments, the pH of the composition is about 6.0 to about 7.5. In some embodiments, the composition is stable at 4°C and / or 25°C for at least 24 hours. In some embodiments, the rapamycin in the nanoparticles has an amorphous form. In some embodiments, the nanoparticle composition is a nanoparticle suspension. In some embodiments, the nanoparticle composition is a dry composition. In some embodiments, the nanoparticle composition is sterilized, for example by filtration. In some embodiments, the nanoparticle composition is contained in a sealed container, for example, a sealed vial or a sealed bag. In some embodiments, the nanoparticle composition contains less than 10 μg / mL of tert-butanol and / or less than 5 μg / mL of chloroform.

[0178] In some embodiments, the nanoparticle composition comprises (a) nanoparticles containing rapamycin and albumin (e.g., human albumin), wherein about 70% to about 85% of the albumin in the nanoparticles is in the form of monomeric albumin, and (b) a non-nanoparticle portion containing albumin (e.g., human albumin) and rapamycin.

[0179] In some embodiments, the nanoparticle composition comprises (a) nanoparticles containing rapamycin and albumin (e.g., human albumin), wherein about 25% to about 50% of the albumin in the nanoparticles is in the form of monomeric albumin, and (b) a non-nanoparticle portion containing albumin (e.g., human albumin) and rapamycin.

[0180] In some embodiments, the nanoparticle composition comprises (a) nanoparticles comprising rapamycin and albumin (e.g., human albumin), wherein about 5% to about 15% of the albumin in the nanoparticles is in the form of high molecular weight albumin (or trimer albumin); and (b) a non-nanoparticle portion comprising albumin (e.g., human albumin) and rapamycin.

[0181] In some embodiments, the nanoparticle composition comprises (a) nanoparticles containing rapamycin and albumin (e.g., human albumin), wherein about 25% to about 50% of the albumin in the nanoparticles is in the form of high molecular weight albumin (other than oligomeric albumin), and (b) a non-nanoparticle portion containing albumin (e.g., human albumin) and rapamycin.

[0182] In some embodiments, the nanoparticle composition comprises (a) nanoparticles containing rapamycin and albumin (e.g., human albumin), wherein about 9% to about 20% of the albumin in the nanoparticles is in the form of dimeric albumin, and (b) a non-nanoparticle portion containing albumin (e.g., human albumin) and rapamycin.

[0183] In some embodiments, the nanoparticle composition comprises (a) nanoparticles containing rapamycin and albumin (e.g., human albumin), wherein about 5% to about 16% of the albumin in the nanoparticles is in the form of dimeric albumin, and (b) a non-nanoparticle portion containing albumin (e.g., human albumin) and rapamycin.

[0184] In some embodiments, the nanoparticle composition comprises (a) nanoparticles comprising rapamycin and albumin (e.g., human albumin), wherein about 70% to about 85% of the albumin in the nanoparticles is in the form of monomeric albumin, about 9% to about 20% of the albumin in the nanoparticles is in the form of dimeric albumin, and about 5% to about 15% of the albumin in the nanoparticles is in the form of polymeric albumin (or trimeric albumin); and (b) a non-nanoparticle portion comprising albumin (e.g., human albumin) and rapamycin.

[0185] In some embodiments, the nanoparticle composition comprises (a) nanoparticles containing rapamycin and albumin (e.g., human albumin), wherein about 25% to about 50% of the albumin in the nanoparticles is in the form of monomeric albumin, about 1% to about 4.5% of the albumin in the nanoparticles is in the form of oligomeric albumin, about 5% to about 16% of the albumin in the nanoparticles is in the form of dimeric albumin, and about 25% to about 50% of the albumin in the nanoparticles is in the form of high molecular weight albumin (other than oligomeric albumin); and (b) a non-nanoparticle portion containing albumin (e.g., human albumin) and rapamycin.

[0186] In some embodiments, the nanoparticle composition comprises (a) nanoparticles having a Z-average particle size of about 200 nm or less (e.g., about 50 nm to about 200 nm), comprising rapamycin and albumin (e.g., human albumin), wherein about 70% to about 85% of the albumin in the nanoparticles is in the form of monomeric albumin, about 9% to about 20% of the albumin in the nanoparticles is in the form of dimeric albumin, and about 5% to about 15% of the albumin in the nanoparticles is in the form of polymeric albumin (or trimeric albumin); and (b) a non-nanoparticle portion comprising albumin (e.g., human albumin) and rapamycin.

[0187] In some embodiments, the nanoparticle composition comprises (a) nanoparticles having a Z-average particle size of about 200 nm or less (e.g., about 50 nm to about 200 nm), comprising a coating containing albumin (e.g., human albumin) and a core containing rapamycin, wherein about 70% to about 85% of the albumin in the nanoparticles is in the form of monomeric albumin, about 9% to about 20% of the albumin in the nanoparticles is in the form of dimeric albumin, and about 5% to about 15% of the albumin in the nanoparticles is in the form of polymeric albumin (or trimeric albumin); and (b) a non-nanoparticle portion containing albumin (e.g., human albumin) and rapamycin.

[0188] In some embodiments, the nanoparticle composition comprises (a) nanoparticles having a Z-average particle size of about 200 nm or less (e.g., about 50 nm to about 200 nm), containing about 55% to about 65% (by weight) of rapamycin and about 25% to about 45% (by weight) of albumin (e.g., human albumin), wherein about 70% to about 85% of the albumin in the nanoparticles is in the form of monomeric albumin, about 9% to about 20% of the albumin in the nanoparticles is in the form of dimeric albumin, and about 5% to about 15% of the albumin in the nanoparticles is in the form of polymeric albumin (or trimeric albumin); and (b) a non-nanoparticle portion containing albumin (e.g., human albumin) and rapamycin.

[0189] In some embodiments, the nanoparticle composition comprises (a) nanoparticles having a Z-average particle size of about 200 nm or less (e.g., about 50 nm to about 200 nm), comprising a coating containing albumin (e.g., human albumin) and a core containing rapamycin, wherein albumin constitutes about 25% to about 45% by weight of the nanoparticles, rapamycin constitutes about 55% to about 75% by weight of the nanoparticles, about 70% to about 85% of the albumin in the nanoparticles is in the form of monomeric albumin, about 9% to about 20% of the albumin in the nanoparticles is in the form of dimeric albumin, and about 5% to about 15% of the albumin in the nanoparticles is in the form of polymeric albumin (or trimeric albumin); and (b) a non-nanoparticle portion containing albumin (e.g., human albumin) and rapamycin.

[0190] In some embodiments, the nanoparticle composition comprises (a) nanoparticles having a Z-average particle size of about 200 nm or less (e.g., about 50 nm to about 200 nm), containing about 55% to about 75% (by weight) of rapamycin and about 25% to about 45% (by weight) of albumin (e.g., human albumin), wherein about 70% to about 85% of the albumin in the nanoparticles is in the form of monomeric albumin, about 9% to about 20% of the albumin in the nanoparticles is in the form of dimeric albumin, and about 5% to about 15% of the albumin in the nanoparticles is in the form of polymeric albumin (or trimeric albumin); and (b) a non-nanoparticle portion containing albumin (e.g., human albumin) and rapamycin, wherein the concentration of rapamycin in the nanoparticle composition is about 1 mg / mL to about 100 mg / mL (e.g., about 1 mg / mL to about 15 mg / mL).

[0191] In some embodiments, the nanoparticle composition comprises (a) a coating containing albumin (e.g., human albumin) and a core containing rapamycin, and is a nanoparticle having a Z-average particle size of about 200 nm or less (e.g., about 50 nm to about 200 nm), wherein the albumin constitutes about 25% to about 45% by weight of the nanoparticle, the rapamycin constitutes about 55% to about 75% by weight of the nanoparticle, and about 70% to about 85% of the albumin in the nanoparticle is in the form of monomeric albumin. The nanoparticle comprises (b) a non-nanoparticle portion containing albumin (e.g., human albumin) and rapamycin, wherein the concentration of rapamycin in the nanoparticle composition is approximately 1 mg / mL to approximately 100 mg / mL (e.g., approximately 1 mg / mL to approximately 15 mg / mL).

[0192] In some embodiments, the nanoparticle composition is (a) nanoparticles having a Z-average particle size of about 200 nm or less (e.g., about 50 nm to about 200 nm) and a zeta potential of about -25 mV to about -50 mV, comprising about 55% to about 75% (by weight) of rapamycin and about 25% to about 45% (by weight) of albumin (e.g., human albumin), wherein about 70% to about 85% of the albumin in the nanoparticles is in the form of monomeric albumin, and the albumin in the nanoparticles The nanoparticles consist of (b) a non-nanoparticle portion comprising albumin (e.g., human albumin) and rapamycin, wherein the concentration of rapamycin in the nanoparticle composition is approximately 1 mg / mL to approximately 100 mg / mL (e.g., approximately 1 mg / mL to approximately 15 mg / mL).

[0193] In some embodiments, the nanoparticle composition comprises (a) a coating containing albumin (e.g., human albumin) and a core containing rapamycin, with a Z-average particle size of about 200 nm or less (e.g., about 50 nm to about 200 nm) and a zeta potential of about 25 mV to about 50 mV, wherein albumin constitutes about 25% to about 45% by weight of the nanoparticles, rapamycin constitutes about 55% to about 75% by weight of the nanoparticles, and about 70% to about 85% of the albumin in the nanoparticles is monomeric albumin. The nanoparticles are in the form of albumin, with approximately 9% to 20% of the albumin in the nanoparticles being in the form of dimeric albumin, and approximately 5% to 15% of the albumin in the nanoparticles being in the form of high molecular weight albumin (or trimer albumin); and (b) a non-nanoparticle portion containing albumin (e.g., human albumin) and rapamycin, wherein the concentration of rapamycin in the nanoparticle composition is approximately 1 mg / mL to 100 mg / mL (e.g., approximately 1 mg / mL to 15 mg / mL).

[0194] In some embodiments, the nanoparticle composition comprises (a) nanoparticles having a Z-average particle size of about 200 nm or less (e.g., about 50 nm to about 200 nm) and a zeta potential of about 25 mV to about 50 mV, comprising about 55% to about 75% (by weight) of rapamycin and about 25% to about 45% (by weight) of albumin (e.g., human albumin), wherein about 70% to about 85% of the albumin in the nanoparticles is in the form of monomeric albumin and about 9% to about 20% of the albumin in the nanoparticles is in the form of dimeric albumin (b) a non-nanoparticle portion comprising (a) a non-nanoparticle portion comprising albumin (e.g., human albumin) and rapamycin, wherein the concentration of rapamycin in the nanoparticle composition is approximately 1 mg / mL to approximately 100 mg / mL (e.g., approximately 1 mg / mL to approximately 15 mg / mL), and approximately 3% or less of the rapamycin in the nanoparticle composition is free rapamycin.

[0195] In some embodiments, the nanoparticle composition comprises (a) a coating containing albumin (e.g., human albumin) and a core containing rapamycin, and is a nanoparticle having a Z-average particle size of about 200 nm or less (e.g., about 50 nm to about 200 nm) and a zeta potential of about 25 mV to about 50 mV, wherein albumin constitutes about 25% to about 45% by weight of the nanoparticle, rapamycin constitutes about 55% to about 75% by weight of the nanoparticle, and about 70% to about 85% of the albumin in the nanoparticle is in the form of monomeric albumin. The nanoparticle comprises (b) a non-nanoparticle portion containing albumin (e.g., human albumin) and rapamycin, wherein the concentration of rapamycin in the nanoparticle composition is approximately 1 mg / mL to approximately 100 mg / mL (e.g., approximately 1 mg / mL to approximately 15 mg / mL), and approximately 3% or less of the rapamycin in the nanoparticle composition is free rapamycin.

[0196] In some embodiments, the nanoparticle composition is a nanoparticle having a Z-average particle size of about 200 nm or less (e.g., about 50 nm to about 200 nm) and a zeta potential of about -25 mV to about -50 mV, comprising (a) about 55% to about 75% (by weight) of rapamycin and about 25% to about 45% (by weight) of albumin (e.g., human albumin), wherein about 70% to about 85% of the albumin in the nanoparticles is in the form of monomeric albumin, and about 9% to about 20% of the albumin in the nanoparticles is in the form of dimeric albumin. The composition comprises (b) nanoparticles in which approximately 5% to approximately 15% of lubumin is in the form of high molecular weight albumin (or trimer albumin), and (b) a non-nanoparticle portion comprising albumin (e.g., human albumin) and rapamycin, wherein the concentration of rapamycin in the nanoparticle composition is approximately 1 mg / mL to approximately 100 mg / mL (e.g., approximately 1 mg / mL to approximately 15 mg / mL), and the total amount of seco-rapamycin and rapamycin in the nanoparticles is less than 3% by weight (e.g., approximately 0.2% to approximately 3%) of seco-rapamycin. In some embodiments, the total amount of seco-rapamycin and rapamycin in the composition is less than 3% by weight (e.g., approximately 0.2% to approximately 3%) of seco-rapamycin.

[0197] In some embodiments, the nanoparticle composition comprises (a) a coating containing albumin (e.g., human albumin) and a core containing rapamycin, and is a nanoparticle having a Z-average particle size of about 200 nm or less (e.g., about 50 nm to about 200 nm) and a zeta potential of about 25 mV to about 50 mV, wherein albumin constitutes about 25% to about 45% by weight of the nanoparticle, rapamycin constitutes about 55% to about 75% by weight of the nanoparticle, about 70% to about 85% of the albumin in the nanoparticle is in the form of monomeric albumin, and about 9% to about 20% of the albumin in the nanoparticle is The present invention comprises (b) a non-nanoparticle portion comprising (a) albumin in the form of dimeric albumin, where about 5% to about 15% of the albumin in the nanoparticles is in the form of high molecular weight albumin (or trimer albumin); and (b) a non-nanoparticle portion comprising albumin (e.g., human albumin) and rapamycin, wherein the concentration of rapamycin in the nanoparticle composition is about 1 mg / mL to about 100 mg / mL (e.g., about 1 mg / mL to about 15 mg / mL), and the total of seco-rapamycin and rapamycin in the nanoparticles is less than 3% by weight (e.g., about 0.2% to about 3%) of seco-rapamycin. In some embodiments, seco-rapamycin constitutes less than 3% (e.g., about 0.2% to about 3%) of the total of seco-rapamycin and rapamycin in the composition.

[0198] In some embodiments, the nanoparticle composition comprises (a) nanoparticles containing rapamycin and albumin (e.g., human albumin), wherein about 74% to about 80% of the albumin in the nanoparticles is in the form of monomeric albumin, and (b) a non-nanoparticle portion containing albumin (e.g., human albumin) and rapamycin. In some embodiments, about 1.5% to about 3% of the albumin in the non-nanoparticle portion or the total albumin in the nanoparticle composition is in the form of high molecular weight albumin (or trimeric albumin). In some embodiments, about 7% to about 11% of the albumin in the non-nanoparticle portion of the nanoparticle composition is in the form of dimeric albumin. In some embodiments, about 7% to about 11% of the total albumin in the nanoparticle composition is in the form of dimeric albumin. In some embodiments, about 83% to about 92% of the albumin in the non-nanoparticle portion or the total albumin in the nanoparticle composition is in the form of monomeric albumin. In some embodiments, the weight ratio of albumin to rapamycin in the composition is about 7:1 to about 9:1. In some embodiments, more than 95% of the albumin in the composition is in the non-nanoparticle portion. In some embodiments, about 98% to about 99.5% of the rapamycin in the composition is in the nanoparticles. In some embodiments, the concentration of albumin in the nanoparticle composition in the non-nanoparticle portion or the total albumin concentration in the nanoparticle composition is about 35 mg / mL to about 45 mg / mL.

[0199] In some embodiments, the nanoparticle composition comprises (a) nanoparticles comprising rapamycin and albumin (e.g., human albumin), wherein about 7% to about 11% of the albumin in the nanoparticles is in the form of high molecular weight albumin (or trimer albumin); and (b) a non-nanoparticle portion comprising albumin (e.g., human albumin) and rapamycin.

[0200] In some embodiments, the nanoparticle composition comprises (a) nanoparticles containing rapamycin and albumin (e.g., human albumin), wherein about 12% to about 17% of the albumin in the nanoparticles is in the form of dimeric albumin, and (b) a non-nanoparticle portion containing albumin (e.g., human albumin) and rapamycin.

[0201] In some embodiments, the nanoparticle composition comprises (a) nanoparticles comprising rapamycin and albumin (e.g., human albumin), wherein about 74% to about 80% of the albumin in the nanoparticles is in the form of monomeric albumin, about 12% to about 17% of the albumin in the nanoparticles is in the form of dimeric albumin, and about 7% to about 11% of the albumin in the nanoparticles is in the form of polymeric albumin (or trimeric albumin); and (b) a non-nanoparticle portion comprising albumin (e.g., human albumin) and rapamycin.

[0202] In some embodiments, the nanoparticle composition comprises (a) nanoparticles having a Z-average particle size of about 85 nm to about 95 nm, comprising rapamycin and albumin (e.g., human albumin), and (b) a non-nanoparticle portion comprising albumin (e.g., human albumin) and rapamycin.

[0203] In some embodiments, the nanoparticle composition comprises (a) nanoparticles having a Z-average particle size of about 85 nm to about 95 nm, comprising rapamycin and albumin (e.g., human albumin), wherein about 74% to about 80% of the albumin in the nanoparticles is in the form of monomeric albumin, about 12% to about 17% of the albumin in the nanoparticles is in the form of dimeric albumin, and about 7% to about 11% of the albumin in the nanoparticles is in the form of polymeric albumin (or trimeric albumin); and (b) a non-nanoparticle portion comprising albumin (e.g., human albumin) and rapamycin.

[0204] In some embodiments, the nanoparticle composition comprises (a) nanoparticles having a Z-average particle size of about 85 nm to about 95 nm, comprising a coating containing albumin (e.g., human albumin) and a core containing rapamycin, wherein about 74% to about 80% of the albumin in the nanoparticles is in the form of monomeric albumin, about 12% to about 17% of the albumin in the nanoparticles is in the form of dimeric albumin, and about 7% to about 11% of the albumin in the nanoparticles is in the form of polymeric albumin (or trimeric albumin); and (b) a non-nanoparticle portion comprising albumin (e.g., human albumin) and rapamycin.

[0205] In some embodiments, the nanoparticle composition comprises (a) nanoparticles having a zeta potential of about -33 mV to about -39 mV, comprising rapamycin and albumin (e.g., human albumin), and (b) a non-nanoparticle portion comprising albumin (e.g., human albumin) and rapamycin.

[0206] In some embodiments, the nanoparticle composition comprises (a) nanoparticles having a zeta potential of about -33 mV to about -39 mV, comprising a coating containing albumin (e.g., human albumin) and a core containing rapamycin, and (b) a non-nanoparticle portion comprising albumin (e.g., human albumin) and rapamycin.

[0207] In some embodiments, the nanoparticle composition comprises (a) nanoparticles having a zeta potential of about -33 mV to about -39 mV, comprising rapamycin and albumin (e.g., human albumin), wherein about 74% to about 80% of the albumin in the nanoparticles is in the form of monomeric albumin, about 12% to about 17% of the albumin in the nanoparticles is in the form of dimeric albumin, and about 7% to about 11% of the albumin in the nanoparticles is in the form of polymeric albumin (or trimeric albumin); and (b) a non-nanoparticle portion comprising albumin (e.g., human albumin) and rapamycin.

[0208] In some embodiments, the nanoparticle composition comprises (a) nanoparticles having a zeta potential of about -33 mV to about -39 mV, comprising a coating containing albumin (e.g., human albumin) and a core containing rapamycin, wherein about 74% to about 80% of the albumin in the nanoparticles is in the form of monomeric albumin, about 12% to about 17% of the albumin in the nanoparticles is in the form of dimeric albumin, and about 7% to about 11% of the albumin in the nanoparticles is in the form of polymeric albumin (or trimeric albumin); and (b) a non-nanoparticle portion comprising albumin (e.g., human albumin) and rapamycin.

[0209] In some embodiments, the nanoparticle composition comprises (a) nanoparticles having a Z-average particle size of about 85 nm to about 95 nm and a zeta potential of about -33 mV to about -39 mV, comprising rapamycin and albumin (e.g., human albumin), and (b) a non-nanoparticle portion comprising albumin (e.g., human albumin) and rapamycin.

[0210] In some embodiments, the nanoparticle composition comprises (a) nanoparticles having a Z-average particle size of about 85 nm to about 95 nm and a zeta potential of about -33 mV to about -39 mV, comprising a coating containing albumin (e.g., human albumin) and a core containing rapamycin, and (b) a non-nanoparticle portion containing albumin (e.g., human albumin) and rapamycin.

[0211] In some embodiments, the nanoparticle composition comprises (a) nanoparticles having a Z-average particle size of about 85 nm to about 95 nm and a zeta potential of about -33 mV to about -39 mV, comprising rapamycin and albumin (e.g., human albumin), wherein about 74% to about 80% of the albumin in the nanoparticles is in the form of monomeric albumin, about 12% to about 17% of the albumin in the nanoparticles is in the form of dimeric albumin, and about 7% to about 11% of the albumin in the nanoparticles is in the form of polymeric albumin (or trimeric albumin); and (b) a non-nanoparticle portion comprising albumin (e.g., human albumin) and rapamycin.

[0212] In some embodiments, the nanoparticle composition comprises (a) nanoparticles having a Z-average particle size of about 85 nm to about 95 nm and a zeta potential of about -33 mV to about -39 mV, comprising a coating containing albumin (e.g., human albumin) and a core containing rapamycin, wherein about 74% to about 80% of the albumin in the nanoparticles is in the form of monomeric albumin, about 12% to about 17% of the albumin in the nanoparticles is in the form of dimeric albumin, and about 7% to about 11% of the albumin in the nanoparticles is in the form of polymeric albumin (or trimeric albumin); and (b) a non-nanoparticle portion comprising albumin (e.g., human albumin) and rapamycin.

[0213] In some embodiments, the nanoparticle composition comprises (a) nanoparticles having a Z-average particle size of about 85 nm to about 95 nm, comprising about 62% to about 68% (by weight) of rapamycin and about 32% to about 38% (by weight) of albumin (e.g., human albumin), wherein about 74% to about 80% of the albumin in the nanoparticles is in the form of monomeric albumin, about 12% to about 17% of the albumin in the nanoparticles is in the form of dimeric albumin, and about 7% to about 11% of the albumin in the nanoparticles is in the form of polymeric albumin (or trimeric albumin); and (b) a non-nanoparticle portion comprising albumin (e.g., human albumin) and rapamycin.

[0214] In some embodiments, the nanoparticle composition comprises (a) nanoparticles having a Z-average particle size of about 85 nm to about 95 nm, comprising about 62% to about 68% (by weight) of rapamycin and about 32% to about 38% (by weight) of albumin (e.g., human albumin), wherein about 74% to about 80% of the albumin in the nanoparticles is in the form of monomeric albumin, about 12% to about 17% of the albumin in the nanoparticles is in the form of dimeric albumin, and about 7% to about 11% of the albumin in the nanoparticles is in the form of polymeric albumin (or trimeric albumin); and (b) a non-nanoparticle portion comprising albumin (e.g., human albumin) and rapamycin, wherein the concentration of rapamycin in the nanoparticle composition is about 1 mg / mL to about 100 mg / mL (e.g., about 1 mg / mL to about 15 mg / mL).

[0215] In some embodiments, the nanoparticle composition comprises (a) nanoparticles having a Z-average particle size of about 85 nm to about 95 nm and a zeta potential of about -33 mV to about -39 mV, comprising about 62% to about 68% (by weight) of rapamycin and about 32% to about 38% (by weight) of albumin (e.g., human albumin), wherein about 74% to about 80% of the albumin in the nanoparticles is in the form of monomeric albumin, and about 12% to about 17% of the albumin in the nanoparticles is The nanoparticles consist of (b) a non-nanoparticle portion comprising albumin (e.g., human albumin) and rapamycin, wherein the concentration of rapamycin in the nanoparticle composition is approximately 1 mg / mL to approximately 100 mg / mL (e.g., approximately 1 mg / mL to approximately 15 mg / mL).

[0216] In some embodiments, the nanoparticle composition comprises (a) nanoparticles having a Z-average particle size of about 85 nm to about 95 nm and a zeta potential of about -33 mV to about -39 mV, comprising about 62% to about 68% (by weight) of rapamycin and about 32% to about 38% (by weight) of albumin (e.g., human albumin), wherein about 74% to about 80% of the albumin in the nanoparticles is in the form of monomeric albumin, and about 12% to about 17% of the albumin in the nanoparticles is in the form of dimeric albumin. (b) Nanoparticles in which approximately 7% to approximately 11% of the albumin in the nanoparticles is in the form of polymer albumin (or trimer albumin); and (b) a non-nanoparticle portion containing albumin (e.g., human albumin) and rapamycin, wherein the concentration of rapamycin in the nanoparticle composition is approximately 1 mg / mL to approximately 100 mg / mL (e.g., approximately 1 mg / mL to approximately 15 mg / mL), and approximately 1% or less of the rapamycin in the nanoparticle composition is free rapamycin.

[0217] In some embodiments, the nanoparticle composition comprises (a) nanoparticles having a Z-average particle size of about 85 nm to about 95 nm and a zeta potential of about -33 mV to about -39 mV, wherein (a) about 62% to about 68% (by weight) of rapamycin and about 32% to about 38% (by weight) of albumin (e.g., human albumin), and about 74% to about 80% of the albumin in the nanoparticles is in the form of monomeric albumin, and about 12% to about 17% of the albumin in the nanoparticles is in the form of dimeric albumin, and about The composition comprises (b) a non-nanoparticle portion comprising (a) nanoparticles in the form of high molecular weight albumin (or trimer albumin) at 7% to approximately 11%, and (b) a non-nanoparticle portion comprising albumin (e.g., human albumin) and rapamycin, wherein the concentration of rapamycin in the nanoparticle composition is approximately 1 mg / mL to approximately 100 mg / mL (e.g., approximately 1 mg / mL to approximately 15 mg / mL), and the total amount of seco-rapamycin and rapamycin in the nanoparticles is less than 1% by weight (e.g., approximately 0.5% to approximately 1%) of seco-rapamycin. In some embodiments, seco-rapamycin is more than approximately 0.2% (e.g., approximately 0.2% to approximately 3%) of the total amount of seco-rapamycin and rapamycin in the composition.

[0218] This specification also provides commercially available batches of nanoparticle compositions (e.g., pharmaceutical compositions) for use in any of the therapeutic methods described herein. As used herein, “commercial batch” refers to a batch size of at least about 20 grams (by mass of rapamycin). Commercial batches are produced on a larger scale than experimental or bench-scale batches. The increase in scale involves longer manufacturing times, including longer processes (such as evaporation processes) or longer waiting times between each process.

[0219] In some embodiments, the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition, e.g., FYARRO®) is administered subcutaneously. In some embodiments, the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition, e.g., FYARRO®) is administered intravenously. In some embodiments, the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition, e.g., FYARRO®) is administered, for example, by intravenous injection at a dose of about 1 mg / m². 2 ~about 150mg / m 2 , about 5mg / m 2 ~about 75mg / m 2 It is administered in the following doses. In some embodiments, the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition, e.g., FYARRO®) is administered, for example, by intravenous injection at doses of about 5, 7.5, 10, 15, 30, 56, 75, or 100 mg / m². 2It is administered in one of the following doses. In some embodiments, the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition, e.g., FYARRO®) is administered to an individual with cancer in one or more 21-day cycles (e.g., a 3-week cycle). In some embodiments, the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition, e.g., FYARRO®) is administered once to the individual during each 21-day cycle (e.g., a 3-week cycle). In some embodiments, the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition, e.g., FYARRO®) is administered in the first, second, or third week of each 21-day cycle (e.g., a 3-week cycle). In some embodiments, the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition, e.g., FYARRO®) is administered on day 1, day 8, or day 15 of each 21-day cycle (e.g., a 3-week cycle). In some embodiments, the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition, e.g., FYARRO®) is administered to the individual twice during each 21-day cycle (e.g., a 3-week cycle). In some embodiments, the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition, e.g., FYARRO®) is administered in the first and second weeks of each 21-day cycle (e.g., a 2-week cycle). In some embodiments, the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition, e.g., FYARRO®) is administered in the second and third weeks of each 21-day cycle (e.g., a 3-week cycle). In some embodiments, the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition, e.g., FYARRO®) is administered in the first and third weeks of each 21-day cycle (e.g., a 3-week cycle). In some embodiments, an mTOR inhibitor nanoparticle composition (e.g., a sirolimus / albumin nanoparticle composition, e.g., FYARRO®) is administered on day 1 and day 8 of each 21-day cycle (e.g., a 3-week cycle).In some embodiments, the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition, e.g., FYARRO®) is administered on day 1 and day 15 of each 21-day cycle (e.g., a 3-week cycle). In some embodiments, the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition, e.g., FYARRO®) is administered on day 8 and day 15 of each 21-day cycle (e.g., a 3-week cycle). In some embodiments, the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition, e.g., FYARRO®) is administered to the individual three times during each 21-day cycle (e.g., a 3-week cycle). In some embodiments, the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition, e.g., FYARRO®) is administered during weeks 1, 2, and 3 of each 21-day cycle (e.g., a 3-week cycle). In some embodiments, the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition, e.g., FYARRO®) is administered on days 1, 8, and 15 of each 21-day cycle (e.g., a 3-week cycle). In some embodiments, the dose of the mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition, e.g., FYARRO®) is modified (e.g., if the individual experiences one or more adverse effects). Details regarding dose modifications for FYARRO® and the circumstances under which dose modifications are made are detailed in www.accessdata.fda.gov / drugsatfda_docs / label / 2021 / 213312lbl.pdf.

[0220] V. Products and Kits In some embodiments, products of manufacture containing materials useful for the treatment by the methods provided herein for undifferentiated pleomorphic sarcoma (including undifferentiated pleomorphic sarcoma with PTEN loss and / or TSC2 mutation) or leiomyosarcoma (including estrogen receptor-positive leiomyosarcoma), etc., and products of manufacture such as drugs or drug combinations comprising an mTOR inhibitor nanoparticle composition (e.g., sirolimus / albumin nanoparticle composition, e.g., nab-sirolimus) and an anti-PD-1 antibody (e.g., nivolumab). The product may include a container and a label or accompanying information on or affixed to the container. Suitable containers include, for example, bottles, vials, syringes, etc. Containers may be formed from various materials such as glass or plastic. Generally, the container may hold a composition effective for the treatment of the disease or disorder described herein and may have a sterile access port (e.g., the container may be an intravenous solution bag or vial with a stopper that can be punctured by a subcutaneous needle). At least one active agent in the composition is either a) a nanoparticle formulation of an mTOR inhibitor, or b) an anti-PD-1 antibody. The label or accompanying information indicates that the composition is used to treat a specific condition in an individual. The label or accompanying information further includes instructions for administering the composition to an individual. Products and kits of manufacture comprising the combination therapies described herein are also contemplated.

[0221] A package insert refers to the instructions typically included with the market packaging of a therapeutic product, containing information regarding the use, dosage, administration, contraindications, and / or warnings for the therapeutic product. In some embodiments, the package insert indicates that the composition is used to treat solid tumors (e.g., bladder cancer, renal cell carcinoma, or melanoma).

[0222] Additionally, the product may further include a second (or third) container containing a pharmaceutically acceptable buffer such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution, and dextrose solution. It may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, and syringes.

[0223] Useful kits are also provided for various purposes, for example, for the treatment of undifferentiated pleomorphic sarcoma (including undifferentiated pleomorphic sarcoma with PTEN loss and / or TSC2 mutation) or leiomyosarcoma (including estrogen receptor-positive leiomyosarcoma). The kit of the present invention comprises one or more containers containing an mTOR inhibitor nanoparticle composition (e.g., a sirolimus / albumin nanoparticle composition) (or a unit dosage form and / or product of manufacture), and in some embodiments further comprises instructions for use with an anti-PD-1 antibody (e.g., one described herein) and / or one of the methods described herein. The kit may further comprise instructions for selecting an individual suitable for treatment. Instructions supplied within the kit of the present invention are typically written instructions on a label or accompanying document (e.g., a paper sheet included in the kit), but machine-readable instructions (e.g., instructions delivered on a magnetic or optical storage disk) are also acceptable.

[0224] The kit of the present invention is contained in suitable packaging. Suitable packaging includes, but is not limited to, vials, bottles, jars, and flexible packaging (e.g., sealed Mylar or plastic bags). The kit may optionally provide additional components such as buffers and explanatory information. Accordingly, the present application also provides products containing vials (such as sealed vials), bottles, jars, flexible packaging, etc.

[0225] Instructions for the use of mTOR inhibitor nanoparticle compositions (e.g., sirolimus / albumin nanoparticle compositions) and anti-PD-1 antibodies generally include information regarding the dose, administration schedule, and route of administration for the intended treatment. Containers may be unit volumes, bulk packages (e.g., multi-dose packages), or secondary unit doses. For example, a kit may be provided containing a dose of the mTOR inhibitor nanoparticle compositions (e.g., sirolimus / albumin nanoparticle compositions) and anti-PD-1 antibodies disclosed herein, sufficient to provide effective treatment to an individual over any of the following long periods, e.g., 1 week, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 2 weeks, 3 weeks, 4 weeks, 6 weeks, 8 weeks, 3 months, 4 months, 5 months, 7 months, 8 months, 9 months, or longer. The kit may also include multiple unit doses of mTOR inhibitor nanoparticle compositions (e.g., sirolimus / albumin nanoparticle compositions) and anti-PD-1 antibodies, as well as instructions for use, packaged in quantities sufficient for storage and use in a pharmacy, e.g., a hospital pharmacy and a compounding pharmacy.

[0226] Those skilled in the art will recognize that several embodiments are possible within the scope and spirit of the present invention. The present invention is described in more detail hereby by reference to the following non-limiting embodiments. The following embodiments further illustrate the present invention, but should not be construed as limiting its scope. [Examples]

[0227] Example 1 This example demonstrates a Phase I / II clinical trial in which nab-sirolimus and nivolumab are administered to individuals with cancer.

[0228] Patients and Methods Test design This was an open-label, single-center, dose-response phase IB trial using a fixed dose of nivolumab, an anti-PD-1 antibody, and an intravenously administered, escalating dose of nab-sirolimus. Patients were enrolled from September 2017 to July 2021. The trial was conducted in accordance with the Declaration of Helsinki and was approved by the Western Laboratory Institutional Review Board (protocol code 20151429 on September 8, 2017) for human-involved trials.

[0229] The primary endpoint was determining the MTD of nab-sirolimus in combination with nivolumab. Secondary endpoints included disease control rates [DCR: complete response (CR) + partial response (PR) + disease stability], objective response rates (CR + PR), and progression-free survival (PFS) and overall survival (OS) as determined by local radiological evaluation using the Criteria for Treatment of Solid Tumors (RECIST) v1.1. Secondary endpoints also included determination of central PFS and central OS.

[0230] For exploratory endpoints, Pearson's correlation coefficient was used to assess the correlation between responses based on immune-related response criteria (irRECIST) and responses based on RECIST v1.1.

[0231] Phase I portion of the study's dose escalation This trial used a standard "3-cohort" design, treating three patients at each dose level, and expanding to six patients per cohort when dose-limiting toxicity (DLT) was observed in one of the three initially enrolled patients at each dose level. If no DLT occurred after two doses, escalation to the next dose level was permitted. MTD was defined as the highest dose at which one or fewer patients experienced DLT and which was safely tolerated, with at least two patients experiencing DLT at the next higher dose level. Patients in the dose-escalation trial could continue treatment at their designated dose levels for up to 18 three-week cycles, or until significant disease progression or unacceptable toxicity occurred. Intra-patient dose escalation did not occur.

[0232] The given dose of nivolumab was 3 mg / kg intravenously for 3 weeks (day 1 of each 21-day cycle), starting from cycle 1. Nab-sirolimus was administered on days 8 and 15, starting from cycle 2. Nivolumab was initially given based on preclinical findings of increased efficacy when administered prior to nab-sirolimus. The escalating dose of nab-sirolimus was 56 mg / m². 2 (n=3~6), 75 mg / m² 2 (n=3-6) and 100 mg / m² 2 Starting with (n=3-6), the drug was administered intravenously for 30 minutes every 3 weeks for 2 weeks.

[0233] It should be noted that DLT includes, according to the Common Terminology Criteria for Adverse Events (CTCAE) v4.03(14), grade 3 or higher colitis, hepatitis, or pneumonia; any grade 1-2 colitis, hepatitis, or pneumonia, adrenal crisis symptoms, any grade 4 hematological toxicity, or any grade 3 or higher non-hematological toxicity that recurs, worsens, or persists with oral steroids for longer than 14 days. DLT is 56 mg / m² 2 If the disease develops in two or more patients, the dose of nab-sirolimus is 45 mg / m² with a DLT of 45 mg / m². 2 If symptoms develop, 45 mg / m² 2 up to, and 30 mg / m² 2 The gradual increase was lifted.

[0234] Expansion phase IB / II portion of the test Following dose escalation and identification of the maximum tolerated dose (MTD), patients continued to receive nab-sirolimus and nivolumab at the MTD, i.e., 100 mg / m2 and 3 mg / kg, respectively. Additional patients added to the study also received nab-sirolimus and nivolumab at the MTD. The objective of the expansion phase was to evaluate overall safety and efficacy in a large number of patients. Patients in the expansion phase of the study continued treatment for up to 18 three-week treatment cycles, or until significant disease progression or unacceptable toxicity occurred. This study was conducted in accordance with the International Council for Harmonisation of Standards for Clinical Trials of Medicinal Products.

[0235] After three or more treatment cycles, the principal investigator recommended surgical debulking, complete surgical resection, or biopsy. If residual disease was present, either by histopathological examination or computerized tomography (CT) scan / magnetic resonance imaging (MRI), the surgical incision healed, and the patient had toxicity less than grade 1, treatment cycles were repeated postoperatively for 2 - 4 weeks.

[0236] Treatment was continued in the presence of an increase in tumor size by CT scan or MRI, when there were no signs or symptoms indicating clear progression, no worsening of the Eastern Cooperative Oncology Group (ECOG) score due to progressive disease, no tumor growth in vital sites threatening life, the patient was aware of alternative therapies and signed an informed consent to delay these therapies, or when there was a clinical benefit determined by the principal investigator, progressive disease was indicated according to RECIST v1.1. Study population

[0237] The study was designed to enroll up to 40 - 50 patients. Patients who did not become evaluable for secondary endpoints by follow-up CT / MRI were replaced. Tumors with the histological structure of Ewing sarcoma, PEComa, epitheloid sarcoma, desmoid tumor, chordoma, non-small cell lung cancer, small cell lung cancer, urothelial carcinoma, melanoma, renal cell carcinoma, head and neck squamous cell carcinoma, hepatocellular carcinoma, classical Hodgkin lymphoma, microsatellite instability / mismatch repair deficiency (MSI-H / dMMR) metastatic colorectal cancer, and tumors with genetic mutations sensitive to mTOR inhibitors were locally confirmed by the facility prior to enrollment.

[0238] Patient eligibility Patients were eligible for inclusion in this study only if all of the following criteria a - j were met: a. A histologically confirmed diagnosis of a tumor with a genetic mutation sensitive to mTOR inhibitors, which is any of Ewing sarcoma, PEComa, epithelioid sarcoma, tendinoid tumor, chordoma, non-small cell lung cancer, small cell lung cancer, urothelial cancer, melanoma, renal cell carcinoma, head and neck squamous cell carcinoma, hepatocellular carcinoma, classical Hodgkin lymphoma, MSIH / dMMR metastatic colorectal cancer, and either metastatic or locally advanced, and for which surgery was not a recommended option. b. One or more measurable target lesions by CT scan or MRI, measurable disease according to RECIST v1.1 confirmed by the investigator. c. Prior therapy was completed at least 5 half-lives or more than 28 days before registration, whichever is shorter. d. Persons aged 12 - 17 years or older, or 18 years or older, with an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1 and weighing ≥ 40 kg. e. Blood chemistry levels at screening (collected within 14 days before registration, local laboratory): total bilirubin ≤ 1.5 × upper limit of normal (ULN) mg / dl, aspartate aminotransferase / alanine aminotransferase ≤ 2.5 × ULN (≤ 5 × ULN if due to liver metastasis), alkaline phosphatase < 3 × ULN (or ≥ 3 × ULN if due to bone metastasis), serum creatinine ≤ 1.5 × ULN. f. Blood cell counts at screening (collected within 14 days before registration, local laboratory): absolute neutrophil count ≥ 1.5 × 10 9 / l, platelet count ≥ 100,000 / mm3 (100 × 10 9 / l), hemoglobin ≥ 9 g / dl, serum triglyceride < 300 mg / dl, serum cholesterol < 350 mg / dl. g. Men and women of reproductive age were required to agree to use effective contraception from 28 days prior to the start of treatment with the study drug, throughout the study period, and after the completion of treatment with the study drug. They were also required to present a negative serum pregnancy test (human chorionic gonadotropin) result at screening and to agree to undergo pregnancy tests during the study period and after the completion of treatment with the study drug. A second form of contraception was also required in the case of tubal ligation. Male patients were required to agree to practice abstinence or use condoms during sexual contact with pregnant or potentially pregnant women while participating in the study. A second form of contraception was also required in the case of successful excision. h. Average life expectancy of 3 months or more as determined by the principal investigator of the clinical trial. i. The ability to understand and provide signed informed consent. j. Willingness and ability to follow scheduled visits, laboratory tests, and other examination procedures.

[0239] Patients were not eligible to be included in this study if any of the following criteria applied: a. Known active, uncontrolled or symptomatic central nervous system (CNS) metastases. In patients with controlled, asymptomatic CNS metastases, prior treatment for CNS metastases (including radiotherapy / surgery) must be completed at least 28 days prior to the initiation of treatment in this study, and chronic corticosteroid therapy for CNS metastases should not be received. b. Active gastrointestinal bleeding, c. Uncontrolled existing thyroid abnormalities, d. A serious, uncontrolled medical or psychiatric disorder. e. Except for patients with non-melanoma skin cancer, cervical cancer, resected incidental prostate cancer (Gleason score ≤6 and postoperative prostate-specific antigen 0.5 ng / ml, graded pT2), or other appropriately treated primary cancers, “active” secondary malignancies, f. If a patient has completed treatment and has been disease-free for more than one year, they are not considered to currently have an active malignant tumor. g. Prior treatment with liver-targeted therapy, radiotherapy [including radiolabeled spheres or CyberKnife, hepatic artery embolization (with or without chemotherapy), or cryotherapy / resection] within two months of enrollment is acceptable if these therapies did not affect the measurable area of ​​disease for which this protocol is used. h. Infections requiring systemic anti-infective treatment completed within 14 days of registration (excluding uncomplicated urinary tract or upper respiratory tract infections), i. Uncontrolled diabetes mellitus, defined by HbA1c ≥ 8%, despite appropriate treatment. j. Unstable coronary artery disease or myocardial infarction in the preceding six months, k. Receiving any combination antitumor therapy. l. A history of interstitial lung disease, pneumonia, or pulmonary hypertension. m. Use potent inhibitors and inducers of cytochrome P450 3A4 (CYP3A4) 14 days prior to receiving the first dose of nab-sirolimus. Additionally, use any known CYP3A4 substrate with a narrow therapeutic window (e.g., fentanyl, alfentanyl, astemizole, cisapride, dihydroergotamine, pimozide, quinidine, terphanide) within 14 days of receiving the first dose of nab-sirolimus. n. Active hepatitis B or hepatitis C, o. Non-oncological vaccine therapy used for the prevention of infection within 4 weeks of trial registration. p. Autoimmune diseases including rheumatoid arthritis, systemic progressive sclerosis (scleroderma), systemic lupus erythematosus, autoimmune vasculitis, and motor neuropathy considered to be autoimmune (e.g., Guillain-Barré syndrome), q. Systemic immunosuppression, including a positive human immunodeficiency virus condition with or without acquired immunodeficiency syndrome. r. Skin rashes affecting 25% or more of the body surface area (psoriasis, eczema), s. Inflammatory bowel disease (Crohn's disease or ulcerative colitis), t. Ongoing or uncontrolled diarrhea within 4 weeks of trial registration. u. Recent history of acute diverticulitis, intra-abdominal abscess, or gastrointestinal obstruction within 6 months of clinical trial registration (known risk factors for intestinal perforation), v. Current, active, or past history of heavy alcohol abuse w. Pituitary secretory disorder, a. Renal failure or excess.

[0240] Exam period The trial was designed to take approximately 32 months from the first patient enrolled to the last patient followed up, including approximately 24 months of enrollment, an estimated 6 months of treatment (or until treatment was no longer permissible), and the end of treatment visits 4 weeks (±7 days) after the last treatment.

[0241] The study was designed to terminate either on the date of the last visit to the last patient, or on the date of receipt of the last data point from the last patient required for primary, secondary, or exploratory analysis, as pre-specified in the protocol.

[0242] The study treatment was designed to end for patients at the date of the final dose of nivolumab or nab-sirolimus. The end of a patient's treatment visit occurred when safety evaluations and procedures were performed after the last treatment, which had to be at least 4 weeks (±7 days) after the final dose of nivolumab or nab-sirolimus.

[0243] The trial was designed with a follow-up period based on the duration of the trial after the completion of treatment visits. All patients who discontinued the study drug and did not give full consent to participate in the trial continued to the follow-up phase for survival and initiation of anticancer therapy. Follow-up was conducted approximately every 12 weeks (±3 weeks) until death, withdrawal of consent, or closure of the trial, whichever came first. This assessment may be performed by record review, with or without telephone contact.

[0244] Statistical considerations Safety analysis: Demographic and baseline information of the trial patients (e.g., extent of previous treatment) was tabulated. The number of patients tested in the dose escalation part of the trial was 9, and 22 for the dose expansion part.

[0245] For the Phase I part of the trial, information on dose level, type (organ effects such as absolute neutrophil count or laboratory determination), severity [by CTCAE version 4.03 for laboratory determination and the most extreme outliers], and relationship to the trial treatment was reported for all observed adverse events. For each dose, the number and percentage of patients who experienced any grade 3, 4, or 5 adverse events, and the number and percentage of patients who experienced selected specific types of adverse events were reported. Further, DLT was summarized by dose level and MTD was determined.

[0246] For the Phase I part of the trial, the full treatment population (full analysis set) was analyzed for all safety analyses. The summary tables provided herein include the number and percentage of patients with adverse events, serious adverse events, fatal adverse events, and other adverse events of interest. Safety was analyzed together in all patient groups (metastatic and locally advanced disease). Outcome data were analyzed using frequency tables, graphs, and summary statistics.

[0247] The incidence of adverse events that occurred during all treatments was tabulated. Tables of fatal adverse events, serious adverse events, treatment-related adverse events, and adverse events leading to discontinuation from the investigational drug were also collected.

[0248] For exposure to nivolumab and nab-sirolimus, summary statistics were used for the total number of doses, the mean dose administered, and the duration of each treatment.

[0249] Efficacy analysis: Disease control rate (CR, PR, SD), objective response rate (ORR), progression-free survival (PFS), and overall survival (OS) were evaluated by local radiological assessment using RECIST v1.1 and irRECIST.

[0250] The focus of the trial was to estimate the disease-recovery rate (DCR) in patients treated with nivolumab and nab-sirolimus. Patients whose disease had progressed before receiving nab-sirolimus were replaced and not included in the statistical analysis. The number and percentage of patients who achieved a response were summarized.

[0251] Other efficacy endpoints, including central PFS and central OS, were analyzed together for all tumor subtypes.

[0252] The number of patients in each category was relatively small; therefore, the median PFS and median OS of these patients were summarized using descriptive statistics.

[0253] Exploratory analysis: Responses based on RECIST v1.1 were correlated with responses based on irRECIST(10) using Pearson's correlation coefficient.

[0254] result Demographic information A total of 34 patients were included in this study. Table 1 shows the patients enrolled according to sex and race. There were 19 males and 15 females, the majority of whom (22 / 34) were Caucasian and not of Hispanic origin. Table 2 shows the patients enrolled according to age group and sex. The majority (56%) were young adults aged 18–39 years. Table 3 shows the histological tumor subtypes of the patients enrolled in the study. The majority of patients (15 / 34) had osteosarcoma or Ewing's sarcoma. [Table 1] [Table 2] [Table 3]

[0255] Overview of Safety Analysis Of the 34 patients who received at least one dose of nab-sirolimus, 31 were eligible for safety analysis.

[0256] No DLTs occurred in Phase I of the trial. In the Phase I portion of the trial, the standard dose of nivolumab was administered every three weeks at days 8 and 15 of each cycle, with doses of 3 mg / kg and 100 mg / m². 2 The MTD dose of nab-sirolimus was used.

[0257] AE (Energetic Exposure) through the Body System Table 4 shows the systemic adverse events (AEs), toxicity grades, and attribution to treatment regimens observed in Phase I and Expansion Phase IB of the study. Of the 31 patients who received at least one dose of nab-sirolimus, 25 (80.6%) had at least one treatment-related AE. Of the 25 patients in the Dose 3+ Expansion Phase IB category, 20 (80%) had at least one treatment-related AE. Grade 3 adverse events thought to be related to nivolumab included increased thyroid-stimulating hormone (3.2%), while grade 3 adverse events thought to be related to nab-sirolimus included thrombocytopenia (9.7%), oral mucositis (3.2%), increased thyroid-stimulating hormone (3.2%), acute dehydration (3.2%), hypertriglyceridemia (3.2%), and hypophosphatemia (3.2%). [Table 4-1] [Table 4-2] [Table 4-3] [Table 4-4] [Table 4-5]

[0258] Summary of effectiveness analysis Table 5 shows the best overall response, PFS, and OS data for patients enrolled in the trial. [Table 5]

[0259] Of the 34 patients who received at least one treatment cycle and a follow-up CT scan, 31 were evaluable for best overall response, PFS, and OS. Three patients did not receive nab-sirolimus.

[0260] The best responders (i.e., those with a partial response) were patients with undifferentiated pleomorphic sarcoma whose tumors had loss of phosphatase tensin homolog (PTEN) and tuberous sclerosis complex 2 (TSC2) mutations, as well as patients with leiomyosarcoma whose tumors were estrogen receptor positive.

[0261] At the data cutoff date, two patients (one with TSC2 and the other with TP53 mutation) were still alive at 113 and 104 weeks, two patients with EWSR1-WT1 fusion were still alive at 104 and 106 weeks, and one patient with PIK3CA was still alive at 98 weeks.

[0262] Summary of exploratory endpoint analysis. Correlation between RECISTv1.1 and irRECIST: Responses were correlated according to RECISTv1.1 and irRECIST using Pearson correlation. The Pearson correlation coefficient was 1.00 (p<0.0001), indicating a perfect correlation.

[0263] Deaths: Table 6 lists deaths, Table 7 lists patients who dropped out due to adverse events, and Table 8 lists patients who were lost to follow-up and therefore their survival status is unknown. As of the data cutoff date, 23 patients had died from disease progression, 8 with disease progression were lost to follow-up, and 5 patients were still alive. Patients did not die within 28 days after receiving nab-sirolimus. [Table 6] [Table 7] [Table 8]

[0264] Table 9 shows the histological structure of each patient's sarcoma, known mutations (multiple mutations possible), number of treatment cycles, best overall response rate (BORR), progression-free survival (PFS), and overall survival (OS). [Table 9-1] [Table 9-2] [Table 9-3] [Table 9-4] [Table 9-5] [Table 9-6] [Table 9-7] [Table 9-8]

[0265] The best responders (PR) were patients with UPS who also had PTEN loss and a TSC2 mutation, as well as patients with LMS who were ER+. Eight patients had a PFS of over 17.8 weeks (4.1 months = median PFS with trabectedin monotherapy). These patients had Ewing's disease (EWSR1 fusion), chondrosarcoma (IDH1 mutation), chondrosarcoma (MSH1, 2, 6, PMS+), and serous carcinoma of the endometrium (P1K3CA mutation or TP53 mutation). One patient with osteosarcoma and one patient with fibrous small cell tumor had no mutations. Many responders had mutations in TSC1 / 2 / mTOR and were low for intermediate TMB and MMR deficiency (dMMR).

[0266] The results demonstrate that treatment with nivolumab plus nab-sirolimus was safe without unexpected adverse events. Unexpectedly, the best responders were found to be patients with anaplastic pleomorphic sarcoma and estrogen receptor-positive leiomyosarcoma with PTEN loss and TSC2 mutations.

Claims

1. A method for treating undifferentiated pleomorphic sarcoma in an individual requiring treatment for undifferentiated pleomorphic sarcoma, wherein the method involves the individual, (a) an effective amount of a composition comprising nanoparticles containing an mTOR inhibitor and albumin, (b) The method comprising administering an effective amount of anti-PD-1 antibody.

2. The method according to claim 1, wherein the undifferentiated pleomorphic sarcoma has a loss of phosphatase tensin homolog (PTEN).

3. The method according to claim 2, wherein the PTEN loss is a loss-of-function mutation or epigenetic silencing.

4. The method according to claim 2 or 3, wherein the individual is selected for the treatment based on having the PTEN loss.

5. The method according to any one of claims 2 to 4, further comprising selecting the individual based on having the aforementioned PTEN loss.

6. The method according to any one of claims 1 to 5, wherein the undifferentiated pleomorphic sarcoma has a tuberous sclerosis complex 2 (TSC2) mutation.

7. The method according to claim 4, wherein the TSC2 mutation is a missense mutation, a nonsense mutation, a deletion, a splicing site mutation, an insertion, a substation, a rearrangement, or a frameshift, or a combination thereof.

8. The method according to claim 6 or 7, wherein the individual is selected for the treatment based on the TSC2 mutation.

9. The method according to any one of claims 6 to 8, further comprising selecting the individual based on having the TSC2 mutation.

10. A method for treating leiomyosarcoma in an individual requiring treatment for leiomyosarcoma, wherein the method involves the individual, (a) an effective amount of a composition comprising nanoparticles containing an mTOR inhibitor and albumin, (b) The method comprising administering an effective amount of anti-PD-1 antibody.

11. The method according to claim 10, wherein the leiomyosarcoma is an estrogen receptor-positive leiomyosarcoma.

12. The method according to claim 11, wherein the individual is selected for the treatment based on having the estrogen receptor-positive leiomyosarcoma.

13. The method according to claim 11 or 12, further comprising selecting the individual based on having the estrogen receptor-positive leiomyosarcoma.

14. The method according to any one of claims 10 to 13, wherein the leiomyosarcoma is PTEN positive.

15. The method according to any one of claims 10 to 14, wherein the leiomyosarcoma has a moderate tumor gene mutation load.

16. The method according to any one of claims 1 to 15, wherein the undifferentiated pleomorphic sarcoma or leiomyosarcoma is locally progressive, progressive, malignant, progressively malignant, or metastatic.

17. The method according to any one of claims 1 to 16, wherein the undifferentiated pleomorphic sarcoma or leiomyosarcoma is recurrent, refractory, or resistant to previous treatment.

18. The method according to claim 17, wherein the aforementioned prior treatment comprises a composition comprising an mTOR inhibitor and albumin-containing nanoparticles.

19. The amount of the mTOR inhibitor in the mTOR inhibitor nanoparticle composition is approximately 10 mg / m ２ ~Approx. 150mg / m ２ The method according to any one of claims 1 to 18.

20. The amount of the mTOR inhibitor in the mTOR inhibitor nanoparticle composition is approximately 100 mg / m ２ The method according to claim 19.

21. The amount of the mTOR inhibitor in the mTOR inhibitor nanoparticle composition is approximately 75 mg / m ２ The method according to claim 19.

22. The amount of the mTOR inhibitor in the mTOR inhibitor nanoparticle composition is approximately 56 mg / m ２ The method according to claim 19.

23. The amount of the mTOR inhibitor in the mTOR inhibitor nanoparticle composition is approximately 45 mg / m ２ The method according to claim 19.

24. The amount of the mTOR inhibitor in the mTOR inhibitor nanoparticle composition is approximately 30 mg / m ２ The method according to claim 19.

25. The method according to any one of claims 1 to 24, wherein the mTOR inhibitor nanoparticle composition is administered weekly.

26. The method according to any one of claims 1 to 25, wherein the mTOR inhibitor nanoparticle composition is administered twice at 3-week intervals.

27. The method according to any one of claims 1 to 26, wherein the mTOR inhibitor nanoparticle composition is administered on the 8th and 15th days of a 21-day cycle.

28. The method according to any one of claims 1 to 27, wherein the mTOR inhibitor nanoparticle composition and the anti-PD-1 antibody are administered to the individual in parallel.

29. The method according to any one of claims 1 to 27, wherein the mTOR inhibitor nanoparticle composition and the anti-PD-1 antibody are administered sequentially to the individual.

30. The method according to any one of claims 1 to 27, wherein the mTOR inhibitor nanoparticle composition and the anti-PD-1 antibody are administered simultaneously to the individual.

31. The method according to any one of claims 1 to 30, wherein the mTOR inhibitor is a limus-based drug.

32. The method according to claim 31, wherein the limus-based drug is sirolimus.

33. The method according to any one of claims 1 to 32, wherein the average diameter of the nanoparticles in the composition is about 150 nm or less.

34. The method according to claim 33, wherein the average diameter of the nanoparticles in the composition is about 120 nm or less.

35. The method according to any one of claims 1 to 34, wherein the weight ratio of albumin to the mTOR inhibitor in the nanoparticle composition is about 9:1 or less.

36. The method according to any one of claims 1 to 35, wherein the nanoparticles include the mTOR inhibitor associated with the albumin.

37. The method according to claim 36, wherein the nanoparticles include the mTOR inhibitor coated with the albumin.

38. The method according to any one of claims 1 to 37, wherein the mTOR inhibitor nanoparticle composition is administered intravenously, intraarterially, intraperitoneally, intravesically, subcutaneously, intrathecally, intrapulmonaryly, intramuscularly, intratracheally, intraocularly, transdermally, or by inhalation.

39. The method according to claim 38, wherein the mTOR inhibitor nanoparticle composition is administered intravenously.

40. The method according to any one of claims 1 to 39, wherein the anti-PD-1 antibody is selected from the group consisting of nivolumab, pembrolizumab, semiprimab, atezolizumab, dostallimab, durvalumab, and avelumab.

41. The method according to any one of claims 1 to 40, wherein the individual is a human.

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