Radiolabelled antibody conjugates and uses thereof

Abstract

The present disclosure provides radiolabeled antibody conjugates as well as methods of making and uses of these conjugates. The radiolabeled antibody conjugate comprises an antibody that specifically binds CD-38, and at least one radionuclide is optionally conjugated to the antibody via a linker. The radionuclide may be a therapeutic radionuclide or a diagnostic radionuclide. Also provided are methods of making the radiolabeled antibody conjugates, and methods of imaging or treating a subject with cancer using the radiolabeled antibody conjugates.

Description

Technical Field

[0001] The present disclosure relates to anti-CD38 antibodies radiolabeled with radionuclides. Such radiolabeled antibody conjugates can be used for the diagnosis and treatment of cancer. Background Art

[0002] The CD38 protein is a bifunctional exonuclease that catalyzes the conversion of NAD+ to cyclic ADP-ribose (cADPR) and the hydrolysis of cADPR to ADP-ribose. CD38 is one of the antigens expressed in malignant plasma cells and is expressed in various hematologic malignancies, including but not limited to multiple myeloma cells, B-cell chronic lymphocytic leukemia cells, and B-cell acute lymphoblastic leukemia cells.

[0003] With the continuous deepening of research into the molecular mechanisms of cell proliferation, tumor growth, and apoptosis, and the revolutionary advances in molecular biology, the diagnosis and treatment of tumors have entered the molecular level. Molecularly targeted drugs for CD38-positive tumors inhibit or kill tumor cells by inhibiting CD38 protein activity or blocking downstream signal transduction pathways. Furthermore, a prerequisite for targeted therapy to benefit patients is the detection of CD38 expression levels in biopsied lesions. Current guidelines, both domestic and international, emphasize that whenever tumor tissue is available, diagnostic agents should be used to pathologically detect not only the primary lesion but also recurrent or metastatic lesions. However, while routinely used clinically, immunohistochemistry and in situ hybridization, while the gold standard for detecting target proteins or genes, are invasive (due to the need for ex vivo tissue biopsy) and difficult to implement (due to the limited size of biopsies required to obtain biopsies for all suspicious lesions, which may not represent the full range of expression in the lesion and can indeed result in false negatives). In contrast, molecular imaging offers the advantages of being non-invasive and capable of visualizing the expression levels of specific targets within lesions.

[0004] Nuclear medicine has inherent advantages as a means of molecular diagnosis and treatment. By selecting appropriate tumor-targeted molecular probes labeled with different functional radionuclides, a highly specific and sensitive in vivo technology for non-invasive diagnosis and treatment of tumors (i.e., radiation killing) can be established. Radionuclide molecular imaging, as a radiotracer method widely used in clinical diagnostic imaging, has advantages such as better tissue penetration, higher safety, and quantitative analysis compared to optical imaging technology, and is very suitable for clinical translation. Positron emission tomography (PET or PET / CT), which has developed rapidly in recent years, has been widely used and highly valued in the diagnosis, staging, efficacy evaluation, and prognosis assessment of most malignant tumors. One of the key features of PET is that it can provide metabolic and functional information (which cannot be obtained by using CT or MRI), making it more sensitive and more suitable for early diagnosis.

[0005] On the other hand, radionuclides for therapeutic use generally emit alpha or beta rays to produce a series of biological effects in diseased tissues caused by these ionizing radiations. In this case, the tumor macromolecular structure, biological activity and tissue properties are destroyed by the direct and indirect effects of the radiation energy, resulting in the loss of growth ability or death of tumor cells, achieving the goal of tumor treatment. Therefore, in preclinical studies and clinical trials, the use of radionuclide-labeled monoclonal antibodies, antigen-binding fragments and peptides, through single photon emission computed tomography (SPECT) or positron emission computed tomography (PET / CT) imaging techniques, can be used to detect the presence and function of specific cell surface receptors. SUMMARY

[0006] In one aspect, the present disclosure relates to anti-CD38 antibodies radiolabeled with a radionuclide (also referred to as "radiolabeled antibody conjugates"). Such radiolabeled antibody conjugates can be used for the diagnosis and treatment of cancer.

[0007] One aspect of the present disclosure relates to a radiolabeled antibody conjugate represented by Formula I:

[0008] Ab-(L-M) p (I)

[0009] wherein,

[0010] Ab is an antibody or antigen-binding fragment thereof that specifically binds to a CD38 protein,

[0011] L is a linker,

[0012] M is a radionuclide, and

[0013] p is an integer from 1 to 10, preferably 2 to 8, preferably 4 to 8, preferably an integer from 3 to 5.

[0014] In certain embodiments, the linker L of the radiolabeled antibody conjugate is represented by Formula II

[0015] Lk-Y-T (II)

[0016] wherein,

[0017] Lk is absent or a coupling group, such as -C(=S)NH-, -C(=O)NH-, -C(=S)-, -C(=O)-, -S-, -CH2C(=O)-, or ,

[0018] Y is a spacer, and

[0019] T is a chelating group.

[0020] In certain embodiments, the radiolabeled antibody conjugate is represented by one of the following structures:

[0021] 、 、 、 、 、 、 or ,

[0022] in,

[0023] Y is independently selected from a bond, a C1-C 20 Alkylene, C2-C 20 Alkenylene, C2-C 20 Alkynylidene, C1-C 20 Heteroalkylene, C3-C 20 Cycloalkylene, C3-C 20 Heterocyclylene (C3-C 20 heterocyclylene), C6-C 20 Arylene or C5-C 20 heteroarylene, keto, amino, thio, -(C=S)-, -O-, or a combination thereof, optionally substituted with one or more halogen, cyano, hydroxy, alkoxy, monocyclic cycloalkyl, monocyclic heterocyclyl,

[0024] T is independently selected from a ligand derived from any of:

[0025] 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA),

[0026] S-2-methyl-1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid,

[0027] N,N''-bis[2-hydroxy-5-(carboxyethyl)benzyl]ethylenediamine-N,N''-diacetic acid (HBED-CC),

[0028] 1,4,7-Triazacyclononane-1,4,7-triacetic acid (NOTA),

[0029] 2-(4,7-bis(carboxymethyl)-1,4,7-triazanonan-1-yl)glutaric acid (NODAGA),

[0030] 2-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododec-1-yl)pentanedioic acid (DOTAGA),

[0031] 1,4,7-Triazacyclononanephosphinic acid (TRAP),

[0032] 1,4,7-Triazacyclononane-1-[methylene(2-carboxyethyl)phosphinic acid]-4,7-bis[methylene(2-hydroxymethyl)phosphinic acid](NOPO),

[0033] 3,6,9,15-tetraazabicyclo[9.3.1.]pentadeca-1(15),11,13-triene-3,6,9-triacetic acid (PCTA),

[0034] N'-{5-[acetyl(hydroxy)amino]pentyl}-N-[5-({4-[(5-aminopentyl)(hydroxy)amino]-4-oxobutanoyl}amino)pentyl]-N-hydroxysuccinamide (DFO),

[0035] Diethylenetriaminepentaacetic acid (DTPA),

[0036] trans-cyclohexyl-diethylenetriaminepentaacetic acid (CHX-DTPA),

[0037] 1-oxa-4,7,10-triazacyclododecane-4,7,10-triacetic acid (oxo-Do3A),

[0038] N x S 4-x (a group of tetracoordinate chelating agents with N atoms (basic amines or non-basic amides) and thiols as donors to stabilize Tc complexes, especially Tc(V)-oxy complexes, such as N4, N2S2, N3S),

[0039] or a combination thereof,

[0040] M is selected from diagnostic radionuclides, therapeutic radionuclides and non-radioactive elements,

[0041] Preferably, the diagnostic radionuclides are independently selected from 68 Ga, 99m Tc, 89 Zr, 43 Sc, 111 In, 45 Ti, 52 Mn, 59 Fe, 64 Cu, 94m Tc, 67 Ga, 71 As、 72 As、 74 As、 82m Rb, 86 Y or a combination thereof,

[0042] Preferably, the therapeutic radionuclides are independently selected from 177 Lu, 90 Y. 153 Sm, 67 Cu, 89 Sr. 166 Ho, 177 Yb, 47 Sc, 186 Re、 188 Re、 212 Bi, 213 Bi, 149 Pm, 212 Pb, 211 At 223 Ra, 225 Ac, 227 Th, 161 Tb or a combination thereof,

[0043] Preferably, the non-radioactive elements are independently selected from Ga, Fe or a combination thereof, and

[0044] Independently selected from covalent bonds, ionic bonds, van der Waals forces, conjugated bonds or combinations thereof.

[0045] In a preferred embodiment, the radiolabeled antibody conjugate is represented by one of the following structures:

[0046] 、 or , preferably .

[0047] In preferred embodiments, Y is independently selected from:

[0048] 、 、 、 、 、 、 、 、 、 、 、 、 、 or a combination thereof,

[0049] wherein R is independently selected from hydrogen or C1-C 10 Alkyl, n is an integer from 0 to 10.

[0050] In a more preferred embodiment, Y is independently selected from:

[0051] 、 、 、 、 or a combination thereof.

[0052] In a more preferred embodiment, T is independently selected from DFO, DOTA, DTPA, CHX-DTPA, or a combination thereof.

[0053] In a more preferred embodiment, Lk-YT is independently selected from any one of the following:

[0054] 1-(4-isothiocyanatophenyl)-3-[6,17-dihydroxy-7,10,18,21-tetraoxy-27-(N-acetylhydroxyamino)-6,11,17,22-tetraazaheptadecanoyl]thiourea (p-SCN-Bn-DFO),

[0055] [(R)-2-amino-3-(4-aminophenyl)propyl]-trans-(S,S)-cyclohexane-1,2-diamine-pentaacetic acid (p-NH2-Bn-CHX-A''-DTPA),

[0056] [(R)-2-amino-3-(4-isothiocyanatophenyl)propyl]-trans-(S,S)-cyclohexane-1,2-diamine-pentaacetic acid (p-SCN-Bn-CHX-A''-DTPA),

[0057] 1-(p-isothiocyanatobenzyl)-3-methyl-DTPA (1B3M),

[0058] Benzyl isothiocyanate-DTPA (SCN-Bz-DTPA),

[0059] 1-(2)-methyl-4-isocyanatobenzyl-DTPA (MX-DTPA),

[0060] Mercaptoacetyl triglycine (MAG3),

[0061] Mercaptoacetyl diglycine (MAG2),

[0062] DOTAGA, NOTA, NODAGA, DOTA and HBED-CC,

[0063] Preferably, Lk-YT is independently selected from any one of the following:

[0064] DOTAGA, NOTA, NODAGA、 DOTA, HBED-CC, p-SCN-Bn-DFO, p-NH2-Bn-CHX-A"-DTPA, p-SCN-Bn-CHX-A"-DTPA,

[0065] Preferably, Lk-YT is independently selected from any one of the following:

[0066] p-SCN-Bn-DFO, p-NH2-Bn-CHX-A"-DTPA, p-SCN-Bn-CHX-A"-DTPA or a combination thereof.

[0067] In a more preferred embodiment, M is independently selected from 99m Tc, 89 Zr, 177 Lu, 225 Ac or a combination thereof.

[0068] In a further preferred embodiment, the radiolabeled antibody conjugate is represented by any one of the following structures:

[0069] 、 、 、 、 、 or .

[0070] In certain embodiments, the Ab is selected from the group consisting of a monoclonal antibody, a single chain antibody, a chimeric antibody, a humanized antibody, and a fully human antibody.

[0071] In a preferred embodiment, the antigen binding fragment is selected from the group consisting of Fab, Fab', F(ab)2, F(ab')2, Fv and ScFv fragments.

[0072] In certain embodiments, Ab comprises an antibody light chain or fragment thereof, wherein the antibody light chain or fragment thereof comprises LCDR1-3, wherein the LCDR1-3 comprises LCDR1, LCDR2, and LCDR3, wherein the LCDR1 comprises the amino acid sequence as shown in SEQ ID NO: 1; the LCDR2 comprises the amino acid sequence as shown in SEQ ID NO: 2; and the LCDR3 comprises the amino acid sequence as shown in SEQ ID NO: 3.

[0073] In preferred embodiments, the light chain of the antibody or fragment thereof comprises a light chain variable region VL, and the light chain variable region VL comprises an amino acid sequence as set forth in SEQ ID NO: 7.

[0074] In more preferred embodiments, the light chain of the antibody or fragment thereof comprises an amino acid sequence as set forth in SEQ ID NO: 17.

[0075] In preferred embodiments, the light chain of the antibody or fragment thereof comprises an amino acid sequence as set forth in any one of SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, and SEQ ID NO: 15.

[0076] In certain embodiments, Ab comprises a heavy chain of an antibody or fragment thereof comprising a HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO: 4; a HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO: 5; and a HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO: 6.

[0077] In preferred embodiments, the heavy chain of the antibody or fragment thereof comprises a heavy chain variable region VH, and the heavy chain variable region VH comprises an amino acid sequence as set forth in SEQ ID NO: 8.

[0078] In more preferred embodiments, the heavy chain of the antibody comprises an amino acid sequence as set forth in SEQ ID NO: 18.

[0079] In preferred embodiments, the heavy chain of the antibody or fragment thereof comprises an amino acid sequence as set forth in any one of SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, and SEQ ID NO: 16.

[0080] In another aspect, the present disclosure provides a pharmaceutical composition comprising the aforementioned radiolabeled antibody conjugate and a pharmaceutically acceptable carrier.

[0081] In another aspect, the present disclosure provides a kit comprising a radionuclide M and a precursor Ab-(Lk-Y-T') p wherein T' is independently selected from any one of:

[0082] 1,4,7,10-tetraazacyclododecane-N,N',N",N"' -tetraacetic acid (DOTA),

[0083] S-2-methyl-1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid,

[0084] N,N''-bis[2-hydroxy-5-(carboxyethyl)benzyl]ethylenediamine-N,N''-diacetic acid (HBED-CC),

[0085] 1,4,7-Triazacyclononane-1,4,7-triacetic acid (NOTA),

[0086] 2-(4,7-bis(carboxymethyl)-1,4,7-triazanonan-1-yl)glutaric acid (NODAGA),

[0087] 2-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododec-1-yl)pentanedioic acid (DOTAGA),

[0088] 1,4,7-Triazacyclononanephosphinic acid (TRAP),

[0089] 1,4,7-Triazacyclononane-1-[methylene(2-carboxyethyl)phosphinic acid]-4,7-bis[methylene(2-hydroxymethyl)phosphinic acid](NOPO),

[0090] 3,6,9,15-tetraazabicyclo[9.3.1.]pentadeca-1(15),11,13-triene-3,6,9-triacetic acid (PCTA),

[0091] N'-{5-[acetyl(hydroxy)amino]pentyl}-N-[5-({4-[(5-aminopentyl)(hydroxy)amino]-4-oxobutanoyl}amino)pentyl]-N-hydroxysuccinamide (DFO),

[0092] Diethylenetriaminepentaacetic acid (DTPA),

[0093] trans-cyclohexyl-diethylenetriaminepentaacetic acid (CHX-DTPA),

[0094] p-Oxa-4,7,10-triazacyclododecane-4,7,10-triacetic acid (oxo-Do3A),

[0095] N x S 4-x (such as N4, N2S2, N3S),

[0096] or a combination thereof;

[0097] Preferably, Lk-YT is independently selected from any one of the following:

[0098] 1-(4-isothiocyanatophenyl)-3-[6, 17-dihydroxy-7, 10, 18, 21-tetraoxyl-27-(N- acetyloxylamino)-6, 11, 17, 22-tetraazahexacosyl]-thiourea (p-SCN-Bn-DFO),

[0099] [(R)-2-amino-3-(4-aminophenyl)propyl]-trans-(S,S)-cyclohexane-1,2-diamine-pentaacetic acid (p-NH2-Bn-CHX-A"-DTPA),

[0100] [(R)-2-amino-3-(4-aminophenyl)propyl]-trans-(S,S)-cyclohexane-1,2-diamine-pentaacetic acid (p-NH2-Bn-CHX-A"-DTPA),

[0101] 1-(p-isothiocyanatobenzyl)-3-methyl-DTPA (1B3M),

[0102] p-isothiocyanatobenzyl-DTPA (SCN-Bz-DTPA),

[0103] 1-(2)-methyl-4-isothiocyanatobenzyl-DTPA (MX-DTPA),

[0104] mercaptoacetyltriglycine (MAG3),

[0105] mercaptoacetyldiglycine (MAG2),

[0106] DOTAGA, NOTA, NODAGA, DOTA, and HBED-CC,

[0107] Preferably, Lk-Y-T is independently selected from any one of: p-SCN-Bn-DFO, p-NH2-Bn-CHX-A"-DTPA, p-SCN-Bn-CHX-A"-DTPA, or a combination thereof.

[0108] In another aspect, the present disclosure provides a pharmaceutical composition comprising the aforementioned radiolabeled antibody conjugate and the aforementioned antibody conjugate.

[0109] In another aspect, the present disclosure relates to use of the aforementioned radiolabeled antibody conjugate, the aforementioned pharmaceutical composition, or the aforementioned kit in the manufacture of a medicament for detecting or aiding in the detection of CD38.

[0110] In another aspect, the present disclosure relates to an imaging method comprising:

[0111] administering the aforementioned radiolabeled antibody conjugate or the aforementioned pharmaceutical composition to an animal;

[0112] and detecting the presence of the radiolabeled antibody conjugate in vivo by imaging.

[0113] In preferred embodiments, the presence of the radiolabeled antibody conjugate is detected by positron emission tomography (PET) or single photon emission computed tomography (SPECT).

[0114] In another aspect, the present disclosure relates to a method of treating cancer, comprising administering the aforementioned radiolabeled antibody conjugate or the aforementioned pharmaceutical composition to a subject.

[0115] In a preferred embodiment, the subject is a CD38 positive subject.

[0116] In preferred embodiments, the cancer is a solid tumor.

[0117] In a more preferred embodiment, the cancer is selected from the group consisting of brain cancer, renal cell carcinoma, ovarian cancer, bladder cancer, prostate cancer, breast cancer, hepatocellular carcinoma, bone cancer, colon cancer, non-small cell lung cancer, head and neck squamous cell carcinoma, colorectal cancer, mesothelioma, B-cell lymphoma, and melanoma.

[0118] In a preferred embodiment, the cancer is a hematological cancer.

[0119] In a more preferred embodiment, the cancer is selected from the group consisting of myeloma, lymphoma and leukemia, preferably, the cancer is selected from the group consisting of acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), chronic myelomonocytic leukemia (CMML), Hodgkin's lymphoma, non-Hodgkin's lymphoma and multiple myeloma. BRIEF DESCRIPTION OF THE DRAWINGS

[0120] Figure 1 According to the present disclosure 89 Schematic representation of the radiolabeling protocol for Zr-DFO-NYM1012.

[0121] Figure 2 show 89 PET / CT images of Zr-DFO-NYM1012 in RPMI8226 and MM.1S tumor-bearing mouse models.

[0122] Figure 3 show 89 PET / CT images of competitive inhibition experiments of Zr-DFO-NYM1012 in RPMI8226 and MM.1S tumor-bearing mouse models.

[0123] Figure 4 show 89PET / CT images of Zr-DFO-NYM1012 in the Raji ectopic mouse model.

[0124] Figure 5 show 89 PET / CT images of Zr-DFO-NYM1012 in the Raji orthotopic mouse model.

[0125] Figure 6a Show Injection 89 Research clinical PET / CT images 24 hours after Zr-DFO-NYM1012. Figure 6b Show Injection 89 Research clinical PET / CT images 72 hours after Zr-DFO-NYM1012.

[0126] Figure 7 According to the present disclosure 177 Schematic diagram of the radiolabeling protocol for Lu-DTPA-NYM1012.

[0127] Figure 8a Shown in 177 Changes in tumor volume of 1MM.1S model mice in the Lu-treated and control groups. Figure 8b Shown in 177 Body weight changes of 1MM.1S model mice in the Lu-treated and control groups. Figure 8c Shown in 177 Changes in tumor volume of RPMI8226 model mice in the Lu-treated and control groups. Figure 8d Shown in 177 Body weight changes of RPMI8226 model mice in the Lu-treated and control groups.

[0128] Figure 9 Shown are the body weight changes of ICR mice.

[0129] Figure 10 A physical map of the construction of the disclosed antibody NYM1012 into a complete antibody expression vector is shown.

[0130] Figure 11 shows 89 HPLC and Radio-iTLC spectra of Zr-DFO-NYM1012. Figure 11a Shows 89 HPLC spectrum of Zr-DFO-NYM1012, indicating the radiochemical purity of the product. Figure 11b The results obtained from Example 4 are shown. 89 HPLC spectrum of Zr-DFO-NYM1012, indicating the chemical purity of the product. Figure 11c The results obtained from Example 4 are shown.89 Radio-iTLC spectrum of Zr-DFO-NYM1012. Detailed Description of the Invention

[0132] definition

[0133] While the following terms are believed to be well understood by those of ordinary skill in the art, the definitions are set forth to facilitate explanation of the presently disclosed subject matter.

[0134] “C1-C 20 "Alkyl" refers to a group of straight-chain or branched saturated hydrocarbon groups having 1 to 20 carbon atoms. "C1-C 10 "Alkyl" refers to a group of straight or branched saturated hydrocarbon groups having 1 to 10 carbon atoms. In some embodiments, C1-C6 alkyl or C1-C4 alkyl is preferred. Examples of C1-C6 alkyl include methyl (C1), ethyl (C2), n-propyl (C3), isopropyl (C3), n-butyl (C4), tert-butyl (C4), sec-butyl (C4), isobutyl (C4), n-pentyl (C5), 3-pentyl (C5), pentyl (C5), neopentyl (C5), 3-methyl-2-butyl (C5), tert-pentyl (C5) and n-hexyl (C6). Optionally, the alkyl group may be substituted with one or more substituents, for example, 1 to 5 substituents, 1 to 3 substituents or 1 substituent. Common abbreviations for alkyl include Me (-CH3), Et (-CH2CH3), iPr (-CH(CH3)2), nPr (-CH2CH2CH3), n-Bu (-CH2CH2CH2CH3) or i-Bu (-CH2CH(CH3)2).

[0135] “C2-C 20 "Alkenyl" refers to a group of straight or branched hydrocarbon groups having 2 to 20 carbon atoms and at least one carbon-carbon double bond. In some embodiments, C2-C 10 C2-C6 alkenyl, C2-C6 alkenyl or C2-C4 alkenyl. Examples of C2-C6 alkenyl include vinyl (C2), 1-propenyl (C3), 2-propenyl (C3), 1-butenyl (C4), 2-butenyl (C4), butadienyl (C4), pentenyl (C5), pentadienyl (C5), hexenyl (C6) and the like. Optionally, alkenyl may be substituted with one or more substituents, for example, 1 to 5 substituents, 1 to 3 substituents or 1 substituent.

[0136] “C2-C 20 "Alkynyl" refers to a straight or branched chain hydrocarbon group having 2 to 20 carbon atoms, at least one carbon-carbon triple bond and optionally one or more carbon-carbon double bonds. In some embodiments, C2-C 10Alkynyl, C2-C6 alkynyl or C2-C4 alkynyl. 2-6 Examples of alkynyl groups include, but are not limited to, ethynyl (C2), 1-propynyl (C3), 2-propynyl (C3), 1-butynyl (C4), 2-butynyl (C4), pentynyl (C5), hexynyl (C6), etc. Optionally, the alkynyl group may be substituted with one or more substituents, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0137] “C1-C 20 "Heteroalkyl" refers to the above-mentioned "C1-C 20 The term "alkyl" refers to a group in which one or more (e.g., 1, 2, 3, or 4) carbon atoms are replaced by heteroatoms (e.g., oxygen, sulfur, nitrogen, boron, silicon, or phosphorus).

[0138] “C3-C 20 "Cycloalkyl" refers to a non-aromatic cyclic hydrocarbon radical having 3 to 20 ring carbon atoms and zero heteroatoms. The term cycloalkyl may contain one or more double bonds or triple bonds as long as it is non-aromatic. In some embodiments, C3-C 10 Cycloalkyl, C3-C8 cycloalkyl, or C3-C6 cycloalkyl are particularly preferred. In some embodiments, C5-C6 cycloalkyl is particularly preferred. Cycloalkyl also includes ring systems in which a cycloalkyl group as described herein is fused to one or more aryl or heteroaryl groups, wherein the point of attachment is on the cycloalkyl ring, and in this case, the number of carbon atoms continues to refer to the number of carbon atoms in the cycloalkyl system. Cycloalkyl groups may be substituted with one or more substituents, for example, with 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0139] “C3-C 20 "Heterocyclyl" refers to a group of a 3-20 membered non-aromatic ring system having ring carbon atoms and 1 to 8 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus and silicon. In heterocyclyl groups containing one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom as long as the bond valence permits. The term heterocyclyl may contain one or more double or triple bonds as long as it is non-aromatic. In some embodiments, C3-C 10 Heterocyclic group, C5-C 10 Heterocyclyl, C5-C6 heterocyclyl. Heterocyclyl also includes ring systems in which the above heterocyclyl is fused to one or more cycloalkyl groups, where the point of attachment is on the cycloalkyl ring, or in which the above heterocyclyl is fused to one or more aryl or heteroaryl groups, where the point of attachment is on the heterocyclyl ring; in this case, the number of ring atoms continues to refer to the number of ring atoms in the heterocyclyl ring system. The heterocyclyl may be substituted with one or more substituents, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0140] “C6-C20 "Aryl" refers to a group of a monocyclic or polycyclic (e.g., bicyclic) 4n+2 aromatic ring system having 6-20 ring carbon atoms and zero heteroatoms. In some embodiments, the aryl group has six ring carbon atoms ("C6 aryl", e.g., phenyl). In some embodiments, the aryl group has ten ring carbon atoms ("C 10 Aryl also includes ring systems in which an aryl ring as described above is fused to one or more cycloalkyl or heterocyclyl groups, and the point of attachment is on the aryl ring, in which case the number of carbon atoms continues to refer to the number of carbon atoms in the aryl ring system. Aryl can be substituted with one or more substituents, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0141] “C5-C 20 "Heteroaryl" refers to a radical of a 5- to 20-membered monocyclic or bicyclic 4n+2 aromatic ring system having ring carbon atoms and 1-8 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen and sulfur. In heteroaryl groups containing one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom as long as the bond valence permits. Heteroaryl bicyclic ring systems can contain one or more heteroatoms in one or both rings. Heteroaryl also includes ring systems in which the above-mentioned heteroaryl ring is fused to one or more cycloalkyl or heterocyclyl groups, and the point of attachment is on the heteroaryl ring. In this case, the number of carbon atoms continues to refer to the number of carbon atoms in the heteroaryl ring system. In some embodiments, C5-C 10 Heteroaryl, C5-C8 heteroaryl or C5-C6 heteroaryl. The heteroaryl group may be substituted with one or more substituents, for example, 1 to 5 substituents, 1 to 3 substituents or 1 substituent.

[0142] “C1-C 20 Alkylene, C2-C 20 Alkenylene, C2-C 20 Alkynylidene, C1-C 20 Heteroalkylene, C3-C 20 Cycloalkylene, C3-C 20 Heterocyclylene, C6-C 20 Arylene or C5-C 20 "Heteroarylene" refers to the above-mentioned C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 Alkynyl, C1-C 20 Heteroalkyl, C3-C 20 Cycloalkyl, C3-C 20 Heterocyclic group, C6-C 20 Aryl or C5-C 20 A divalent group formed by removing another hydrogen from a heteroaryl group.

[0143] "Amino" means a -NR2 or -N(R)- group, wherein R refers to H, C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 Alkynyl, C1-C 20 Heteroalkyl, C3-C 20 Cycloalkyl, C3-C 20 Heterocyclic group, C6-C 20 Aryl or C5-C 20 "Alkoxy" refers to -OR, where R is a C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 Alkynyl, C1-C 20 Heteroalkyl, C3-C 20 Cycloalkyl, C3-C 20 Heterocyclic group, C6-C 20 Aryl or C5-C 20 Heteroaryl.

[0144] "Halo" or "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br) and iodine (I).

[0145] "Keto" refers to -(C=O)-. "Thio" refers to -S-. "Cyano" refers to -CN. "Hydroxyl" refers to -OH.

[0146] As used herein, the term "antibody" refers to a peptide that can specifically recognize and / or neutralize a specific antigen. For example, an antibody may comprise an immunoglobulin composed of at least two heavy (H) chains and two light (L) chains connected to each other by disulfide bonds, and may comprise any molecule containing its antigen-binding fragment. The term "antibody" includes monoclonal antibodies, antibody fragments or antibody derivatives, including but not limited to human antibodies (fully human antibodies), humanized antibodies, chimeric antibodies, single-chain antibodies (e.g., scFv) and antigen-binding fragments of antibodies (e.g., Fab, Fab' and (Fab)2 fragments). The term "antibody" also includes all recombinant forms of antibodies, such as antibodies expressed in prokaryotic cells, non-glycosylated antibodies and any antigen-binding fragments of antibodies of the present disclosure and derivatives thereof. Each heavy chain can be composed of a heavy chain variable region (VH) and a heavy chain constant region. Each light chain can be composed of a light chain variable region (VL) and a light chain constant region. The VH and VL regions can be further divided into hypervariable regions (called complementarity determining regions (CDRs, including LCDRs and HCDRs)), which are interspersed between more conserved regions called framework regions (FRs). Each VH and VL can be composed of three CDRs and four FRs, arranged in the following order from amino terminus to carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4. The variable regions of the heavy and light chains contain binding domains that interact with antigens. The constant region of the antibody can mediate the binding of the immunoglobulin to host tissues or factors, including multiple cells of the immune system (such as effector cells) and the first component (Clq) of the classical complement system.

[0147] The term "antigen-binding fragment" refers to one or more fragments of an antibody that specifically binds to an antigen. The antigen-binding function of an antibody can be achieved by a full-length fragment of the antibody. The antigen-binding function of an antibody can also be achieved by a heavy chain comprising an Fv, ScFv, dsFv, Fab, Fab', or F(ab')2 fragment, or a light chain comprising an Fv, ScFv, dsFv, Fab, Fab', or F(ab')2 fragment. (1) Fab fragments, i.e., monovalent fragments comprising the VL, VH, CL, and CH domains; (2) F(ab')2 fragments, bivalent fragments comprising two Fab fragments linked by a disulfide bond at the hinge region; (3) Fd fragments comprising the VH and CH domains; (4) Fv fragments comprising the VL and VH domains in one arm of the antibody; (5) dAb fragments comprising the VH domain (Ward et al., (1989) Nature 341:544-546); (6) isolated complementarity determining regions (CDRs); and (7) combinations of two or more isolated CDRs, optionally linked by a linker. Furthermore, monovalent single-chain molecules Fv (scFv) formed by pairing VL and VH can also be included (see Bird et al., (1988) Science 242: 423-426; and Huston et al., (1988) Proc. Natl. Acad. Sci. 85: 5879-5883).

[0148] As used herein, the term "specifically binds" refers to an antibody that is capable of binding to an antigen of interest (e.g., CD38) with sufficient binding affinity such that the antibody can be used to target the antigen of interest. As an example, the binding affinity of an antibody for the antigen of interest, K D Can be about 10 -4 to about 10 -15 , or about 10 -6 to about 10 -13 , or about 10 -7 to about 10 -12 , or about 10 -9 to about 10 -10 As another example, where the antibody is one that binds CD38, it will preferentially bind to CD38 over other antigens and / or extracellular components. As another example, where the antibody is one that binds CD38, it may not significantly cross-react with other non-CD38 antigens. In embodiments, the extent of binding of the antibody to non-CD38 antigens and / or other substances is less than about 10% (such as 0% to about 9%), as determined by standard techniques known to those of ordinary skill in the art (e.g., by flow cytometric analysis).

[0149] As used herein, the term "CD38" includes any variants, isoforms, and species homologs of CD38 that are naturally expressed in cells or expressed in cells transfected with the CD38 gene. In the present disclosure, CD38 may be human CD38 with GenBank accession number BAA18966.1. In the present disclosure, CD38 may be monkey CD38, such as macaque CD38 with GenBank accession number AAT36330.1. The CD38 protein of the present disclosure may also be referred to as ADP-ribosyl cyclase 1, cADPr hydrolase 1, Cd38-rs1, cyclic ADP-ribose hydrolase 1, or NIM-R5 antigen. "CD38 + " or "CD38 positive" generally refers to cells or subjects that express CD38 protein. It can also be called CD38 positive cells. "CD38 - " or "CD38 negative" generally refers to cells or subjects that do not substantially express CD38 protein.

[0150] As used herein, a "radiolabeled antibody conjugate" is an antibody labeled with a radionuclide. A radiolabeled antibody conjugate is considered to be a combination of an antibody and a radionuclide.

[0151] As used herein, the term "radionuclide" refers to an atom with an unstable nucleus, which is a nucleus with excess energy characteristics that can be transferred to newly generated radiation particles or atomic electrons within the nucleus. In this process, the radionuclide undergoes radioactive decay. Radionuclides can exist naturally, but can also be artificially produced. Radionuclides vary according to their characteristics (including half-life, energy emission characteristics, and decay type). This allows people to select radionuclides with a desired combination of characteristics that are suitable for diagnostic and / or therapeutic uses. For example, gamma emitters are typically used for diagnosis, and beta emitters are typically used for treatment. However, some radionuclides are both gamma emitters and beta emitters, and therefore can be adapted for both uses by changing the amount of radioactivity used (total activity and / or specific activity).

[0152] As used herein, the term "diagnostic radionuclide" refers to a radionuclide moiety that can be attached to an antibody and used to provide a detectable signal. Diagnostic radionuclides include radioisotopes that can be detected by virtue of radioactive decay. Diagnostic radionuclides can be detected by various imaging techniques such as, for example, PET and SPECT imaging. Examples of diagnostic radionuclides suitable for PET or SPECT imaging and suitable for inclusion in labeled antibody conjugates according to embodiments of the present disclosure include 68 Ga, 99m Tc, 89 Zr, 43 Sc, 111In, 45 Ti, 52 Mn, 59 Fe, 64 Cu, 94m Tc, 67 Ga, 71 As、 72 As、 74 As、 82m Rb, 86 Y or a combination of any two or more thereof.

[0153] As used herein, the term "therapeutic radionuclide" refers to a radionuclide moiety that can be attached to an antibody and whose function is to deliver a cytotoxic dose of radiation (e.g., a radionuclide therapeutic agent) to a target of interest (e.g., a tumor). Suitable radionuclide therapeutic agents include radionuclides that emit beta particle radiation, alpha particle radiation, Auger electron radiation, or a combination of any two or more thereof. Examples of suitable radionuclide therapeutic agents include 177 Lu, 90 Y. 153 Sm, 67 Cu, 89 Sr. 166 Ho, 177 Yb, 47 Sc, 186 Re、 188 Re、 212 Bi, 213 Bi, 149 Pm, 212 Pb, 211 At 223 Ra, 225 Ac, 227 Th, 161 Tb or a combination of any two or more thereof.

[0154] The present disclosure contemplates that the antibodies or antigen-binding fragments can be radionuclide labeled using any available methods and chemistries.The binding or conjugation of the radionuclide can be direct binding or conjugation or binding or conjugation via a linker.

[0155] As used herein, the term "linker" refers to a chemical moiety that connects an antibody to a radionuclide label. The linker can include 1) a chelating group for binding the radionuclide label, 2) a coupling group that binds the linker to the antibody or antigen-binding fragment, and 3) optionally, a spacer that connects the chelating group and the coupling group. The linker acts by complexing the radionuclide with the chelating group and covalently linking the chelating group (and the complexed radionuclide) to the antibody through reaction of the coupling group with the corresponding reactive functional group of the antibody.

[0156] Antibody conjugates

[0157] In one or more embodiments, the present disclosure relates to radiolabeled anti-CD38 antibody conjugates.In embodiments, the radiolabeled anti-CD38 antibody conjugates include both diagnostic radionuclide antibody conjugates and therapeutic radionuclide antibody conjugates.

[0158] In one embodiment, the diagnostic radionuclide antibody conjugate comprises an antibody that specifically binds CD-38 and at least one diagnostic radionuclide is conjugated to the antibody, optionally via a linker.

[0159] In one embodiment, the therapeutic radionuclide antibody conjugate comprises an antibody that specifically binds CD38 and at least one therapeutic radionuclide is conjugated to the antibody.

[0160] It has been unexpectedly discovered that by conjugating a radionuclide to an antibody that specifically binds CD38 ("anti-CD38 antibodies"), high radiochemical purity and radiolabeling yield can be achieved.

[0161] In one aspect, good imaging results are achieved by combining diagnostic radionuclides with anti-CD38 antibodies, wherein high intratumoral radiotracer uptake and low background are observed. The resulting radiolabeled antibody conjugates exhibit high tumor selectivity, which is beneficial for observing tumor lesions and can be used for non-invasive monitoring of CD38 protein expression in lesions throughout the body. Further, the resulting radiolabeled antibody conjugates also exhibit desired safety. For example, a PET / CT scan or SPECT / CT scan can be performed on the first day after the patient is injected with the radiolabeled antibody conjugate, so that clinicians can obtain data (including tumor size, location, malignancy and other data) through the imaging results and determine surgical indications and surgical plans, while the radioactivity in the patient disappears by radioactive decay over the next few days (depending on the half-life of the radioisotope used).

[0162] On the other hand, by utilizing a targeted delivery system (e.g., an anti-CD38 antibody), therapeutic radionuclides can be specifically delivered to tumors and accumulated in tumors, thereby inhibiting tumor growth and exhibiting superior anti-tumor effects, indicating excellent radiotherapy potential. Further, the antibody conjugates described herein also exhibit less accumulation in normal tissues, thereby reducing biological toxicity and achieving good safety.

[0163] Furthermore, the molecular structure of the linker disclosed herein exhibits a strong affinity for the targeted antigen CD38 and specifically binds to CD38 on the cell surface, thereby resulting in highly effective ADCC activity against the specifically bound cells.

[0164] Antibody

[0165] The antibodies or antigen-binding fragments thereof according to the present disclosure were measured at 1×10 -9 M or lower K D value (e.g., not greater than about 1×10 -9 M, not more than about 9×10 -10 M, not more than about 8×10 -10 M, not more than about 7×10 -10 M, not more than about 6×10 -10 M, not more than about 5×10 -10 M, not more than about 4×10 -10 M, not more than about 3×10 -10 M, not more than about 2×10 -10 M, not more than about 1×10 -10 M or not greater than about 1×10 -11 M or lower K D value) binds to CD38 protein.

[0166] According to the present disclosure, the antibodies or antigen-binding fragments thereof or variants thereof can precisely target tumor cells and / or inhibit tumor growth by specifically binding to the CD38 protein. For example, the tumor may include a CD38-positive tumor. For example, the CD38-positive tumor may be selected from the group including multiple myeloma, lymphoma, and leukemia. Alternatively, for example, the tumor is selected from the group including non-Hodgkin's lymphoma and Hodgkin's lymphoma. The tumor cells may be selected from the group including Raji cells, Daudi cells, Ramos cells, RPMI8226 cells, and MM.1S cells. In the present disclosure, the antibodies or antigen-binding fragments thereof can kill and damage multiple myeloma, lymphoma, leukemia, non-Hodgkin's lymphoma, and Hodgkin's lymphoma cells or inhibit the growth of multiple myeloma, lymphoma, leukemia, non-Hodgkin's lymphoma, and Hodgkin's lymphoma.

[0167] The antibodies according to the present disclosure may be monoclonal antibodies, single-chain antibodies, chimeric antibodies, humanized antibodies and / or fully human antibodies. The antigen-binding fragments of the present disclosure may be Fab, Fab', F(ab)2, F(ab')2, Fv and / or ScFv fragments.

[0168] Suitable antibodies that specifically bind to CD38 (anti-CD38 antibodies) can be prepared by standard techniques known to those of ordinary skill in the art or obtained commercially.

[0169] The antibody or antigen-binding fragment thereof according to the present disclosure may comprise a light chain of an antibody or a fragment thereof.

[0170] For example, the light chain of the antibody or a fragment thereof may comprise LCDR1, and LCDR1 may comprise the amino acid sequence set forth in SEQ ID NO: 1. The light chain of the antibody or a fragment thereof may comprise LCDR2, and LCDR2 may comprise the amino acid sequence set forth in SEQ ID NO: 2. The light chain of the antibody or a fragment thereof may comprise LCDR3, and LCDR3 may comprise the amino acid sequence set forth in SEQ ID NO: 3.

[0171] The light chain of the antibody of the present disclosure or a fragment thereof may comprise a light chain variable region VL, and the light chain variable region VL may comprise the amino acid sequence shown in SEQ ID NO: 7.

[0172] In the present disclosure, the light chain of the antibody or a fragment thereof may comprise the amino acid sequence shown in SEQ ID NO: 17: EIVMTQSPASLSASLGQRAX 20 ISCRASX 27 SVSX 31 SAX 34 SYVHWYQQKSGQPPKLLIYLASX 57 X 58 X 59 SGVPARFSGSGSGTTDFTLTIIPVESEDVATYYCHHSRX 97 X 98 PX 100 X 101 FGSGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 17), where X 20 =T or S; X 27 =S or N; X 31 =S or T; X 34 =F or Y; X 57 =N and D; X 58 =L or I; X 59 =E or Q; X 97 =E or Q; X 98 =L or V; X 100 =F or S; X 101 =T or S.

[0173] The light chain of the antibody of the present disclosure or a fragment thereof comprises the amino acid sequence shown in any one of SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13 and SEQ ID NO: 15.

[0174] The antibody or antigen-binding fragment thereof according to the present disclosure may comprise a heavy chain of an antibody or a fragment thereof.

[0175] In the present disclosure, the heavy chain of the antibody or a fragment thereof may comprise HCDR1, wherein HCDR1 may comprise the amino acid sequence shown in SEQ ID NO: 4. The heavy chain of the antibody or a fragment thereof may comprise HCDR2, and HCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 5. Alternatively, for example, the heavy chain of the antibody or a fragment thereof may comprise HCDR3, and HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 6.

[0176] The heavy chain of the antibody or a fragment thereof may include a heavy chain variable region VH, and the heavy chain variable region VH may include the amino acid sequence shown in SEQ ID NO: 8.

[0177] In the present disclosure, the heavy chain of the antibody or its fragment may comprise the amino acid sequence shown in SEQ ID NO: 18: QVQLLESGGGLVQPGGSLKLSCVASGX 27 X 28 FSLYX 33 MNWVRQAPGKGLEWIGKIX 52 PX 54 SSX 57 X 58 X 59 YX 61 PSX 6 4 KDKFFISRDNAKNTLYLQMTKVRSEDTALYYCARLX 100 IX 102 X 103 GGX 106 X 107YWGQGTTLTVSSASTKGPSVFPLAPSSKSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 18), where X 27 =F or Y; X 28 =D or N; X 33 =W or Y; X 52 =N, Q or S; X 54 =D, E or N; X 57 =T or S; X 58 =I or L; X 59 =N or Q; X 6 =T or S; X 64 =L or V; X 100 =W or Y; X 102 =A or G; X 103 =T or S; X 106 =F or Y; X 107 =D or N.

[0178] The heavy chain of the antibody of the present disclosure or a fragment thereof comprises the amino acid sequence shown in any one of SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14 and SEQ ID NO: 16.

[0179] In some embodiments, in the antibody or antigen-binding fragment thereof of the present disclosure, LCDR1 can comprise the amino acid sequence as set forth in SEQ ID NO: 1 or a variant thereof, LCDR2 can comprise the amino acid sequence as set forth in SEQ ID NO: 2 or a variant thereof, LCDR3 can comprise the amino acid sequence as set forth in SEQ ID NO: 3 or a variant thereof, HCDR1 can comprise the amino acid sequence as set forth in SEQ ID NO: 4 or a variant thereof, HCDR2 can comprise the amino acid sequence as set forth in SEQ ID NO: 5 or a variant thereof, and HCDR3 can comprise the amino acid sequence as set forth in SEQ ID NO: 6 or a variant thereof. In some embodiments, the light chain or antigen-binding fragment thereof of the antibody of the present disclosure can comprise a light chain variable region, which can comprise the amino acid sequence as set forth in SEQ ID NO: 7 or a variant thereof, and a heavy chain, which can comprise a heavy chain variable region, which can comprise the amino acid sequence as set forth in SEQ ID NO: 8 or a variant thereof. In some embodiments, the antibody or antigen-binding fragment thereof of the present disclosure can comprise a light chain, which can comprise the amino acid sequence as set forth in SEQ ID NO: 9, and a heavy chain, which can comprise the amino acid sequence as set forth in SEQ ID NO: 10.

[0180] In some embodiments, the antibody of the present disclosure can be NYM1012. LCDR1, LCDR2, and LCDR3 of antibody NYM1012 have the amino acid sequences as set forth in SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, respectively; VL has the amino acid sequence as set forth in SEQ ID NO: 7; HCDR1, HCDR2, and HCDR3 have the amino acid sequences as set forth in SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, respectively; VH has the amino acid sequence as set forth in SEQ ID NO: 8; the light chain has the amino acid sequence as set forth in SEQ ID NO: 11; and the heavy chain has the amino acid sequence as set forth in SEQ ID NO: 13.

[0181] The antibodies or antigen-binding fragments thereof disclosed herein may further comprise one or more random mutations (e.g., one or more, one or several amino acid substitutions) in the amino acid sequences of their light and / or heavy chains. For example, the antibodies or antigen-binding fragments thereof may comprise one or more random mutations (e.g., one or more, one or several amino acid substitutions) at one or more sites in the framework regions L-FR1 to L-FR4 of their light chain variable regions, and / or comprise one or more random mutations (e.g., one or more, one or several amino acid substitutions) at one or more sites in the framework regions H-FR1 to H-FR4 of their heavy chain variable regions. For example, upon random mutation, the light chain or antigen-binding fragment thereof of the antibody may comprise an amino acid sequence as set forth in any one of SEQ ID NO: 11, SEQ ID NO: 13, and SEQ ID NO: 15; and / or, the heavy chain or antigen-binding fragment thereof of the antibody may comprise an amino acid sequence as set forth in any one of SEQ ID NO: 12, SEQ ID NO: 14, and SEQ ID NO: 16. The randomly mutated CD38 antibody or antigen-binding fragment thereof still has the ability to specifically bind to human CD38 protein and monkey CD38 protein.

[0182] In some embodiments, the light chain or antigen-binding fragment thereof of the antibody of the present disclosure may comprise the amino acid sequence as shown in SEQ ID NO: 11; and the heavy chain may comprise the amino acid sequence as shown in SEQ ID NO: 12; alternatively, the light chain or antigen-binding fragment thereof of the antibody of the present disclosure may comprise the amino acid sequence as shown in SEQ ID NO: 13; and the heavy chain may comprise the amino acid sequence as shown in SEQ ID NO: 14; alternatively, the light chain or antigen-binding fragment thereof of the antibody of the present disclosure may comprise the amino acid sequence as shown in SEQ ID NO: 15; and the heavy chain may comprise the amino acid sequence as shown in SEQ ID NO: 16.

[0183] The proteins, peptides and / or amino acid sequences involved in the present disclosure should also be understood to cover the following scope: variants or homologs that have the same or similar functions as the proteins or peptides.

[0184] In the present disclosure, a variant can be a protein or peptide obtained by substituting, deleting or adding one or more amino acids relative to the amino acid sequence of a protein and / or peptide (e.g., an antibody or fragment thereof that specifically binds to a CD38 protein). For example, a functional variant can comprise a protein or peptide having an amino acid modification by substitution, deletion and / or insertion of at least one (e.g., 1-30, 1-20, or 1-10, or for example, 1, 2, 3, 4, or 5 amino acids). A functional variant can substantially retain the biological activity of the protein or peptide prior to modification (e.g., substitution, deletion, or addition). For example, a functional variant can retain at least 60%, 70%, 80%, 90%, or 100% of the biological activity (such as antigen binding ability) of the original protein or peptide. For example, the substitution can be a conservative substitution.

[0185] In the present disclosure, a homolog can be a protein or peptide that has at least about 85% (e.g., at least about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or more) sequence identity with the amino acid sequence of the protein and / or peptide (e.g., an antibody or fragment thereof that specifically binds to a CD38 protein).

[0186] In the present disclosure, homology generally refers to the similarity (similarity), similarity (analogy) or relatedness (association) between two or more sequences." sequence homology percentage ratio " can be calculated as follows: Two sequences to be compared are relatively in a comparison window, to determine that there is the number of the position of identical nucleic acid base (for example, A, T, C, G, I) or identical amino acid residue (for example, Ala, Pro, Ser, Thr, Gly, Val, Leu, Ile, Phe, Tyr, Trp, Lys, Arg, His, Asp, Glu, Asn, Gln, Cys and Met) in two sequences, to obtain the number of matching positions.The quantity of matching positions is divided by the sum (i.e. window size) of the position in the comparison window, and the result is multiplied by 100 to produce sequence homology percentage ratio.Comparison for determining sequence homology percentage ratio can be carried out according to various methods known in the art, such as using publicly available computer software, such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithm required for achieving maximum alignment within the full-length sequence being compared or within the target sequence region. Homology can also be determined by the following methods: FASTA and BLAST. For a description of the FASTA algorithm, reference can be made to "Improved Tools for Biological Sequence Comparison" by WRPearson and DJ Lipman, Proceedings of the National Academy of Sciences (Proc. Natl. Acad. Sci.), 85:2444-2448, 1988; and "Fast and Sensitive Protein Mass Similarity Search" by DJ Lipman and WR Pearson, Science, 227:1435-1441, 1989. For a description of the BLAST algorithm, see S. Altschul, W. Gish, W. Miller, E.W. Myers, and D. Lipman, “A Basic Local Alignment Search Tool”, Journal of Molecular Biology, 215:403-410, 1990.

[0187] Conjugation

[0188] In the radiolabeled antibody conjugates according to the present disclosure, a diagnostic radionuclide or a radionuclide therapeutic agent is conjugated directly or indirectly to an anti-CD38 antibody or an anti-CD38 antigen-binding fragment to produce a diagnostic radionuclide antibody conjugate or a therapeutic radionuclide antibody conjugate according to embodiments of the present disclosure.

[0189] In an exemplary embodiment, conjugation of a diagnostic radionuclide or therapeutic radionuclide to an anti-CD38 antibody or anti-CD38 antigen-binding fragment comprises reacting a coupling group of a linker with a corresponding reactive functional group (such as a terminal amino group, a terminal carboxyl group, or a functional group of an amino acid side chain) of the anti-CD38 antibody or anti-CD38 antigen-binding fragment, thereby covalently binding the linker to the antibody or anti-CD38 antigen-binding fragment and complexing the diagnostic radionuclide or therapeutic radionuclide to the linker.

[0190] Non-limiting examples of coupling groups that react with sulfhydryl groups of cysteine-containing antibodies or antigen-binding fragments include epoxides, haloacetyl groups, sulfhydryl groups, and maleimides. Non-limiting examples of coupling groups that react with amino groups of antibodies or antigen-binding fragments include N-hydroxysuccinimide esters, carbodiimides, aldehydes, ketones, glyoxal, iminoesters, isothiocyanates, sulfonyl chlorides, and acyl azides. Non-limiting examples of coupling groups that react with carboxylic acid groups of antibodies or antigen-binding fragments include amines, hydrazides, carbodiimides, diazoalkanes, diazoacetyl groups, and carbonyldiimidazoles.

[0191] For conjugation of a diagnostic or therapeutic radionuclide to an antibody or antigen-binding fragment, a suitable linker can be used, wherein the linker binds to both the antibody or antigen-binding fragment and the diagnostic or therapeutic radionuclide. In certain embodiments, the linker is bifunctional and thus has the function of binding to both the antibody or antigen-binding fragment and the diagnostic or therapeutic radionuclide, resulting in a diagnostic radionuclide antibody conjugate or a therapeutic radionuclide antibody conjugate.

[0192] In exemplary embodiments, the linker comprises a chelating group optionally connected to a coupling group via a spacer.

[0193] Non-limiting examples of chelating groups and chelating groups having spacers and coupling groups include ligands derived from any of the following:

[0194] 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA),

[0195] S-2-methyl-1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid,

[0196] N,N''-bis[2-hydroxy-5-(carboxyethyl)benzyl]ethylenediamine-N,N''-diacetic acid (HBED-CC),

[0197] 1,4,7-Triazacyclononane-1,4,7-triacetic acid (NOTA),

[0198] 2-(4,7-bis(carboxymethyl)-1,4,7-triazanonan-1-yl)glutaric acid (NODAGA),

[0199] 2-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododec-1-yl)pentanedioic acid (DOTAGA),

[0200] 1,4,7-Triazacyclononanephosphinic acid (TRAP),

[0201] 1,4,7-Triazacyclononane-1-[methylene(2-carboxyethyl)phosphinic acid]-4,7-bis[methylene(2-hydroxymethyl)phosphinic acid](NOPO),

[0202] 3,6,9,15-tetraazabicyclo[9.3.1.]pentadeca-1(15),11,13-triene-3,6,9-triacetic acid (PCTA),

[0203] N'-{5-[acetyl(hydroxy)amino]pentyl}-N-[5-({4-[(5-aminopentyl)(hydroxy)amino]-4-oxobutanoyl}amino)pentyl]-N-hydroxysuccinamide (DFO),

[0204] 1-(4-isothiocyanatophenyl)-3-[6,17-dihydroxy-7,10,18,21-tetraoxy-27-(N-acetylhydroxyamino)-6,11,17,22-tetraazaheptadecanoyl]thiourea (p-SCN-Bn-DFO),

[0205] Diethylenetriaminepentaacetic acid (DTPA),

[0206] trans-cyclohexyl-diethylenetriaminepentaacetic acid (CHX-DTPA),

[0207] [(R)-2-amino-3-(4-aminophenyl)propyl]-trans-(S,S)-cyclohexane-1,2-diamine-pentaacetic acid (p-NH2-Bn-CHX-A''-DTPA),

[0208] [(R)-2-amino-3-(4-isothiocyanatophenyl)propyl]-trans-(S,S)-cyclohexane-1,2-diamine-pentaacetic acid (p-SCN-Bn-CHX-A''-DTPA),

[0209] 1-(p-isothiocyanatobenzyl)-3-methyl-DTPA (1B3M),

[0210] 1-oxa-4,7,10-triazacyclododecane-4,7,10-triacetic acid (oxo-Do3A),

[0211] Benzyl isothiocyanate-DTPA (SCN-Bz-DTPA),

[0212] 1-(2)-methyl-4-isocyanatobenzyl-DTPA (MX-DTPA),

[0213] Mercaptoacetyl triglycine (MAG3),

[0214] Mercaptoacetyl diglycine (MAG2),

[0215] N x S 4-x (e.g. N4, N2S2, N3S),

[0216] Non-limiting examples of spacers include: bonds, C1-C 20 Alkylene, C2-C 20 Alkenylene, C2-C 20 Alkynylidene, C1-C 20 Heteroalkylene, C3-C 20 Cycloalkylene, C3-C 20 Heterocyclylene, C6-C 20 Arylene or C5-C 20 Heteroarylene, keto, amino, thio, -(C=S)-, -O-, or a combination thereof, optionally substituted with one or more halogen, cyano, hydroxy, alkoxy, monocyclic cycloalkyl, monocyclic heterocyclyl.

[0217] In an exemplary embodiment, the radiolabeled antibody conjugate is not limited in the number of radionuclides included. In an exemplary embodiment, the radiolabeled antibody conjugate comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more radionuclides. Where multiple labels are included, each label can be the same or different.

[0218] In illustrative examples, the diagnostic radionuclides are independently selected from 68 Ga, 99m Tc, 89 Zr, 43 Sc, 111 In, 45 Ti, 52 Mn, 59 Fe, 64 Cu, 94m Tc,67 Ga, 71 As、 72 As、 74 As、 82m Rb, 86 Y or a combination thereof.

[0219] In an illustrative embodiment, the therapeutic radionuclides are independently selected from 177 Lu, 90 Y. 153 Sm, 67 Cu, 89 Sr. 166 Ho, 177 Yb, 47 Sc, 186 Re、 188 Re、 212 Bi, 213 Bi, 149 Pm, 212 Pb, 211 At 223 Ra, 225 Ac, 227 Th, 161 Tb or a combination thereof.

[0220] In an exemplary embodiment, the linker is attached to the antibody or antibody fragment.In a further embodiment, the radionuclide is complexed to a linker, wherein the linker is covalently attached to the antibody or antibody fragment.

[0221] In an exemplary embodiment, the linker is attached to the antibody or antibody fragment by reacting the coupling group of the linker with the corresponding reactive functional group of the antibody or antibody fragment by standard techniques known to those of ordinary skill in the art.

[0222] In an exemplary embodiment, the ion of the radionuclide tracer is complexed to the linker by standard techniques known to those of ordinary skill in the art.

[0223] In an illustrative, non-limiting embodiment, (1) the linker Df-Bz-NCS is reacted with an anti-CD38 antibody or antibody fragment to covalently attach it; and (2) the radionuclide tracer 89 Zr 4+ The radiolabeled antibody conjugate is formed by reaction with the linker Df-Bz-NCS, which is covalently attached to the antibody or antibody fragment.

[0224] In an illustrative, non-limiting embodiment, (1) the linker p-SCN-Bn-CHX-A''-DTPA is reacted with an anti-CD38 antibody or antibody fragment to covalently attach it; and (2) the radionuclide tracer 177 Lu3+ Reacts with the linker p-SCN-Bn-CHX-A''-DTPA, which is covalently attached to the antibody or antibody fragment, to form a radiolabeled antibody conjugate.

[0225] Pharmaceutical composition

[0226] The pharmaceutical composition according to aspects of the present disclosure comprises a radiolabeled antibody conjugate in an amount ranging from about 0.1-99% and a pharmaceutically acceptable carrier.

[0227] Pharmaceutical compositions of the present disclosure can be any dosage form suitable for application to a subject, illustratively including solid, semisolid and liquid dosage forms, such as tablets, capsules, powders, granules, pills, solutions, suspensions and gels. Liposomes and emulsions are pharmaceutical preparations of the well-known type that can be used for delivering compositions of the present disclosure. Pharmaceutical compositions of the present disclosure generally include a pharmaceutically acceptable carrier, such as an excipient, a diluent and / or a vehicle. Delayed release formulations and delayed release systems of the compositions can be used, such as a semipermeable matrix of a solid hydrophobic polymer.

[0228] The term "pharmaceutically acceptable carrier" refers to a carrier that is suitable for use in a subject without undue toxicity or irritation to the subject and is compatible with the other ingredients included in the pharmaceutical composition.

[0229] Pharmaceutically acceptable carriers, methods for preparing pharmaceutical compositions and various dosage forms, and modes of administration are well known in the art, as described in detail, for example, in LV Allen, Jr. et al., Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems, 8th Ed., Philadelphia, Pa.: Lippincott, Williams & Wilkins, 2004; AR Gennaro, Remington: The Science and Practice of Pharmacy, Lippincott Williams & Wilkins, 21st ed., 2005; and JG Hardman et al., Goodman & Gilman's The Pharmacological Basis of Therapeutics, McGraw-Hill Professional, 10th ed., 2001.

[0230] Solid dosage forms for suspension in a liquid prior to administration illustratively include capsules, tablets, powders, and granules. In such solid dosage forms, one or more active agents are mixed with at least one carrier, which illustratively includes a buffer such as, for example, sodium citrate or an alkali metal phosphate, which illustratively includes sodium phosphate, potassium phosphate, and calcium phosphate; a filler such as, for example, starch, lactose, sucrose, glucose, mannitol, and silicic acid; a binder such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia; a humectant such as, for example, glycerol; a disintegrant such as, for example, agar, calcium carbonate, a plant starch such as potato or Tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; solution retarders such as, for example, paraffin wax; absorption accelerators such as, for example, quaternary ammonium compounds; wetting agents such as, for example, cetyl alcohol, glyceryl monostearate, and ethylene glycol; adsorbents such as, for example, kaolin and bentonite; lubricants such as, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycol, or sodium lauryl sulfate; preservatives such as, for example, antibacterial and antifungal agents, including, for example, sorbic acid, gentamicin, and phenol; and stabilizers such as, for example, sucrose, EDTA, EGTA, and antioxidants.

[0231] Compositions for parenteral administration can be formulated into injectable liquids. Liquid dosage forms for injection include one or more active agents and a pharmaceutically acceptable carrier formulated as an emulsion, solution, or suspension. Liquid dosage forms for injection comprising compositions of the present disclosure may further include stabilizers, wetting agents, emulsifiers, suspending agents, or any two or more thereof. Examples of suitable aqueous and non-aqueous pharmaceutically acceptable carriers include water, buffers, ethanol, polyols such as propylene glycol, polyethylene glycol, glycerol, etc., and suitable mixtures thereof; vegetable oils such as olive oil; and injectable organic esters such as ethyl oleate. For example, by using a coating such as lecithin, by maintaining the desired particle size in the case of a dispersion, and / or by using a surfactant such as sodium lauryl sulfate, appropriate fluidity can be maintained. Optionally, stabilizers such as, for example, sucrose, EDTA, EGTA, and antioxidants are included.

[0232] Imaging and / or treatment approaches

[0233] In one or more embodiments, methods for imaging are disclosed herein. In embodiments, the methods for imaging comprise: (a) administering a radiolabeled antibody conjugate to a subject, wherein the radiolabeled antibody conjugate comprises: an antibody that specifically recognizes and binds to CD38, and at least one diagnostic radionuclide conjugated to the antibody; and (b) detecting the presence of the radiolabeled antibody conjugate in the subject by imaging. In embodiments, the radiolabeled antibody conjugate is as described herein. In further embodiments, the antibody and at least one diagnostic radionuclide are as described herein for antibody conjugates.

[0234] In an exemplary embodiment, the method for imaging comprises detecting the presence of a radiolabeled antibody conjugate in a subject by an imaging technique. In an embodiment, the presence of the radiolabeled antibody conjugate is detected in real time. In an embodiment, the presence of the radiolabeled antibody conjugate is detected non-invasively and / or minimally invasively.

[0235] According to aspects of the present disclosure, the radiolabeled antibody conjugate is an imaging agent that can be used to visualize CD38, such as in diagnostic procedures and for locating tumors or lesions that produce CD38. Imaging can be performed by any of a number of procedures well known to those of ordinary skill in the art, such as positron emission tomography (PET), single photon emission computed tomography (SPECT), Cerenkov imaging, photoacoustic imaging, ultrasound imaging, optical coherence tomography, optical imaging (including fluorescence imaging, magnetic resonance imaging, magnetic particle imaging, bioluminescence imaging), or a combination of any two or more thereof.

[0236] In one or more embodiments, methods for treatment are disclosed herein. In embodiments, the methods for treatment comprise administering to a subject a radiolabeled antibody conjugate, wherein the radiolabeled antibody conjugate comprises: an antibody that specifically recognizes and binds to CD38, and at least one therapeutic radionuclide conjugated to the antibody. In embodiments, the radiolabeled antibody conjugate is as described herein. In further embodiments, the antibody and at least one therapeutic radionuclide are as described herein for antibody conjugates.

[0237] In exemplary embodiments, the methods disclosed herein include administering a radiolabeled antibody conjugate to a subject. In embodiments, the method includes administering an effective amount of a radiolabeled antibody conjugate to a subject. The radiolabeled antibody conjugate can be administered by any suitable route known to those of ordinary skill in the art. In embodiments, the radiolabeled antibody conjugate is administered intravenously, intraarterially, intraperitoneally, intramuscularly, subcutaneously, intrapleurally, intrathecally, intratumorally, by local catheter perfusion and / or direct intralesional injection. In embodiments in which the radiolabeled antibody conjugate is administered by injection, administration can be by continuous infusion, by single bolus injection and / or by multiple bolus injections. In some embodiments, administration of the radiolabeled antibody conjugate is non-immunogenic to the subject. According to exemplary embodiments, administering the radiolabeled antibody conjugate to a subject includes administering a pharmaceutical composition comprising the radiolabeled antibody conjugate and a pharmaceutically acceptable carrier.

[0238] In an exemplary embodiment, the subject is a mammal. In some embodiments, the subject is a mammal selected from the group consisting of humans, non-human primates, canines, felines, murines, bovines, equines, caprines, ovines, porcines, and lagomorphs. According to specific embodiments, the subject is a rodent, including but not limited to rats, mice, or guinea pigs. According to specific embodiments, the subject is a mouse or a human.

[0239] According to an exemplary embodiment, a method for imaging a subject comprises 1) administering to the subject a radiolabeled antibody conjugate, wherein the radiolabeled antibody conjugate comprises: an antibody that specifically binds CD38 and a diagnostic radionuclide conjugated to the antibody; and 2) detecting the presence of the labeled antibody conjugate in the subject by imaging.

[0240] According to an exemplary embodiment, a method for treating a subject comprises administering to the subject a radiolabeled antibody conjugate, wherein the radiolabeled antibody conjugate comprises: an antibody that specifically binds CD38 and a therapeutic radionuclide conjugated to the antibody.

[0241] According to specific embodiments, the subject suffers from cancer. According to specific embodiments, the cancer is a solid tumor. According to specific embodiments, the cancer is brain cancer, renal cell carcinoma, ovarian cancer, bladder cancer, prostate cancer, breast cancer, hepatocellular carcinoma, bone cancer, colon cancer, non-small cell lung cancer, head and neck squamous cell carcinoma, colorectal cancer, mesothelioma, B cell lymphoma and melanoma. According to specific embodiments, the cancer is a hematological cancer. According to specific embodiments, the cancer is myeloma, lymphoma and leukemia, preferably, the cancer is selected from the group consisting of acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), chronic myelomonocytic leukemia (CMML), Hodgkin's lymphoma, non-Hodgkin's lymphoma and multiple myeloma. Example

[0242] The present disclosure is further described below by way of examples, but the present disclosure is not limited to the scope of the examples described. The experimental methods without specific conditions in the following examples are selected according to conventional methods and conditions, or according to the product instructions.

[0243] Example 1: Preparation of hybridoma antibodies and gene cloning

[0244] Mouse Immunization: Balb / c mice (purchased from Beijing Weitonglihua Laboratory Animal Technology Co., Ltd.) were immunized subcutaneously (sc) three times with 100 μg of soluble CD38 antigen (purchased from Beijing Sino-Biotech Co., Ltd.). On day 0, Freund's complete adjuvant was mixed with the antigen, and on days 14 and 28, Freund's incomplete adjuvant was used. Splenocytes from immunized mice were fused with mouse myeloma SP2 / 0 cells (ATCC) according to standard protocols.

[0245] Hybridoma fusion: Mouse spleens were fused according to the current conventional hybridoma fusion method, and the fused hybridoma cells were screened according to the HAT screening method (10 5 cells / well). After 12 days, the supernatant was assayed by ELISA using a microplate coated with CD38 antigen. Selected clones underwent a second round of subcloning by limiting dilution analysis. The resulting hybridoma cell lines stably expressing CD38 antibodies were used for seed storage and library construction.

[0246] RNA preparation: RNA was prepared using the TRIzol RNA extraction kit (Life Technologies). cDNA encoding the antibody gene was prepared using a reverse transcription kit (Beijing Quanshijin Biotechnology Co., Ltd.) and used as a template for PCR amplification of the antibody variable region genes. The cloned heavy and light chain variable region sequences were sequenced to obtain the corresponding amino acid sequences.

[0247] Example 2: Preparation of NYM1012

[0248] Humanization is performed to obtain a humanized antibody sequence.

[0249] The humanized antibody was named NYM1012. Sequencing results showed that the amino acid sequences of LCDR1-3 of antibody NYM1012 were shown in SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, respectively; the amino acid sequence of VL was shown in SEQ ID NO: 7; the nucleotide sequence encoding VL obtained by codon optimization and reverse translation was shown in SEQ ID NO: 19; the amino acid sequences of HCDR1-3 of antibody NYM1012 were shown in SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, respectively; the amino acid sequence of VH was shown in SEQ ID NO: 8; and the nucleotide sequence encoding VH obtained by codon optimization and reverse translation was shown in SEQ ID NO: 20.

[0250] The amino acid sequence of the light chain of antibody NYM1012 is shown in SEQ ID NO: 9, and the nucleotide sequence encoding the same is shown in SEQ ID NO: 21. The amino acid sequence of the heavy chain of antibody NYM1012 is shown in SEQ ID NO: 10, and the nucleotide sequence encoding the same is shown in SEQ ID NO: 22.

[0251] The obtained humanized antibody variable region gene was cloned into the eukaryotic expression vector pCMV-163 containing the human IgG constant region gene to construct a humanized antibody with Figure 10 The complete antibody expression vector with the physical map shown. Figure 10 In the present invention, the various components of the eukaryotic expression vector pCMV-163 are known in the art and are recombined in the order shown.

[0252] The obtained eukaryotic expression vector encoding the antibody NYM1012 was transfected into CHO-S cells using the ExpiCHO™ Expression System Kit (purchased from Thermo Fisher Scientific) for expression, and the cell culture supernatant containing the protein of interest was collected. The target antibody was purified by conventional protein A affinity purification.

[0253] NYM1012 can also be prepared according to the method disclosed in US11713357B2, in particular the preparation of SG003 as shown in Examples 1 and 2 thereof.

[0254] Example 3: Preparation of DFO-NYM1012

[0255]

[0256] DFO-NYM1012

[0257] p-SCN-Bn-DFO (1-(4-isothiocyanatophenyl)-3-[6,17-dihydroxy-7,10,18,21-tetraoxy-27-(N-acetylhydroxyamino)-6,11,17,22-tetraazaheptacosine]thiourea) was purchased directly from Macrocyclics (CAS No.: 1222468-90-7).

[0258] The antibody NYM1012 prepared in Example 2 was dissolved in a 0.1 mol / L NaHCO 3 solution having a pH of 8.4 to achieve an antibody concentration of 10 mg / mL.

[0259] The pH of the resulting antibody solution was adjusted to 9.0 using a 0.1 mol / L Na2CO3 solution. A DMSO solution containing 2 mmol / L p-benzyldeferoxamine isothiocyanate (p-SCN-Bn-DFO) was added to the antibody solution, resulting in a molar ratio of p-SCN-Bn-DFO to the NYM1012 antibody of 5:1. After mixing, the reaction mixture was incubated at 37°C for 60 min to obtain crude DFO-NYM1012.

[0260] The crude DFO-NYM1012 was purified using a PD10 column. After purification, physiological saline and a 3.5 mg / mL gentisic acid solution were added as stabilizers, and the mixture was stored at -80°C and thawed for subsequent use.

[0261] The labeling process was completed within 120 min, and the labeling yield was approximately 50%.

[0262] Embodiment 4: 89 Preparation of Zr-DFO-NYM1012

[0263] The synthesized DFO-NYM1012 (precursor for radiolabeling) was reacted with the radionuclide zirconium-89 [ 89 Zr] chelation to obtain tracers for clinical PET / CT imaging 89 Zr-DFO-NYM1012.

[0264] 100 μL of 0.1 mol / L sodium carbonate solution (pH 10) 89 Zr oxalic acid solution (3 mCi) was neutralized to pH 7.0.

[0265] To the above solution, 0.3 mL of 0.15 mol / L acetic acid-sodium acetate buffer at pH 7.2 was added, followed by the addition of DFO-NYM1012 (2 mg in acetic acid-sodium acetate solution at pH 7.2) and mixed thoroughly to form a reaction mixture. The mixture was allowed to react at room temperature for 60 min, with shaking every 10 minutes. The crude labeled product was thus obtained. 89 Zr-DFO-NYM1012.

[0266] The crude labeled product obtained as described above 89 Zr-DFO-NYM1012 (if the total volume is less than 2.5 mL, the above-mentioned acetic acid-sodium acetate buffer should be added to reach a total volume of 2.5 mL) was added to a PD10 column that had been pre-equilibrated with 0.15 mol / L acetic acid-sodium acetate buffer (pH 7.2) and all pre-equilibration effluent was discarded. 3 mL of the above-mentioned acetic acid-sodium acetate buffer was added to the column, all fractions were collected, and then the combined fractions were filtered through a sterile 0.22 μm filter membrane to obtain the radiolabeled immunoconjugate. 89 Zr-DFO-NYM1012.

[0267] Example 5: 89 Quality Control of Zr-DFO-NYM1012

[0268] In this study, Radio-iTLC and high performance liquid chromatography were used to characterize the radiolabeled products. 89 Zr-DFO-NYM1012 was quality controlled by Radio-iTLC method. 89 Radiochemical purity of Zr-DFO-NYM1012. The supporting material of Radio-iTLC is glass fiber paper, and the developing solvent is 0.5 M citrate / sodium citrate buffer (pH=5). The radiolabeled product sample is pipetted 89Zr-DFO-NYM1012 was taken from its container and spotted lightly on a glass fiber paper about 1.5 cm from the bottom. Then, the glass fiber paper was put into a test tube containing citric acid / sodium citrate buffer. The development was to the solvent front about 2.5 cm from the top of the TLC paper. The glass fiber paper was taken out of the test tube and dried, and further detected by Radio-TLC scanner. In this system, the Rf value of free radionuclide (Zr salt) was between 0.8 and 1.0, while the Rf value of radiolabeled product was between 0 and 0.2. The results showed that the radiochemical purity of Zr-DFO-NYM1012 was 100% (as shown in 89 89 Figure 11c

[0269] The chemical and radiochemical purity of Zr-DFO-NYM1012 was also determined by SEC-HPLC with a mobile phase of 0.1 M phosphate buffer (PB) + 0.2 M NaCl + 10% aqueous acetonitrile. A TSKgel G3000SWXL column was used with a flow rate of 0.8 mL / min and a detection wavelength of 280 nm. It was found that the chemical purity of Zr-DFO-NYM1012 was higher than 95% (as shown in 89 89 Figure 11b Figure 11a 89 The specific activity of radiolabeled Zr-DFO-NYM1012 was found to be about 1 mCi / mg, not more than 1.5 mCi / mg.

[0270] Example 6: 89 Tissue distribution and targeting of Zr-DFO-NYM1012 in RPMI8226 and MM.1S mouse models

[0271] Mouse models: The experimental RPMI8226 model and MM.1S model were provided by Constant Biological Technology (Suzhou) Co., Ltd., which are subcutaneous ectopic RPMI8226 tumor and MM.1S tumor models based on BALB / c nude mice. They are mouse models of human multiple myeloma peripheral blood B lymphocytes and human IgA-myeloma cells, respectively.

[0272] Experimental procedure: 100 mCi of Zr-DFO-NYM1012 prepared as described above was administered to each of two RPMI8226 and two MM.1S mice. 89 ​​​​​​​Zr-DFO-NYM1012. MicroPET / CT imaging scans were performed 24h, 48h, and 96h after administration.

[0273] Before microPET / CT imaging, mice were preanesthetized with an appropriate concentration of isoflurane and air mixture and placed in a microPET / CT imaging chamber (SNPC-303 Super Nova, Pingsheng Medical Technology (Kunshan) Co., Ltd.), where anesthesia was maintained with an isoflurane / air mixture. Scanned images were reconstructed using the scanner software and analyzed using PMOD software.

[0274] Conclusion: Figure 2 As shown, 89 The biodistribution of Zr-DFO-NYM1012 in RPMI8226 and MM.1S mice revealed high radioactive uptake in tumor tissues (indicated by arrows) and low uptake in other tissues, indicating that the radiolabeled product 89 Zr-DFO-NYM1012 has good tumor targeting properties. Furthermore, after administration, high uptake at the tumor site and low uptake in normal tissues result in a high tumor-to-background ratio, which produces high-contrast images at the tumor site and facilitates tumor diagnosis.

[0275] Example 7: 89 Competitive inhibition of Zr-DFO-NYM1012 in RPMI8226 and MM.1S mouse models

[0276] Mouse Models: The RPMI8226 and MM.1S models used in these experiments were provided by Hengjia Biotechnology (Suzhou) Co., Ltd. These are subcutaneous ectopic RPMI8226 and MM.1S tumor models established in BALB / c nude mice. These are mouse models derived from human multiple myeloma peripheral blood B lymphocytes and human IgA-myeloma cells, respectively.

[0277] Experimental procedures: Two RPMI8226 and two MM.1S mice were each given unlabeled NYM1012 antibody via the tail vein. The injection mass of unlabeled NYM1012 antibody was 89 The mass of Zr-DFO-NYM1012 is about 40 times that of unlabeled NYM1012. 89 Zr-DFO-NYM1012 (100 μCi). Before scanning, mice were pre-anesthetized with an appropriate concentration of isoflurane / air mixture and placed in a MicroPET / CT imaging chamber (SNPC-303 Super Nova, Pingsheng Medical Technology (Kunshan) Co., Ltd.), and anesthesia was maintained with isoflurane / air mixture. 89MicroPET / CT scans were performed 24, 48, and 96 hours after Zr-DFO-NYM1012 administration. Scanned images were reconstructed using the scanner software and analyzed using PMOD software.

[0278] Conclusion: Figure 3 As shown, compared with the example 6 without pre-injection of unlabeled NYM1012 antibody Figure 2 In contrast, radioactivity 89 The uptake of Zr-DFO-NYM1012 in tumor tissue was significantly reduced (the arrows indicate the tumor location). This means that the pre-injection of unlabeled NYM1012 significantly blocked the 89 Tumor uptake of Zr-DFO-NYM1012, thus demonstrating 89 Tumor uptake of Zr-DFO-NYM1012 is specific.

[0279] Example 8: 89 Tissue distribution and targeting experiments as well as competitive inhibition studies of Zr-DFO-NYM1012 in the Raji ectopic mouse model

[0280] Mouse Model: The Raji mouse model used in the experiment was provided by Biocytogen (Beijing) Pharmaceutical Technology Co., Ltd. It is an ectopic model based on the subcutaneous transplantation of Raji tumors in B-NDG mice. This model is a mouse tumor model established by using human Burkitt's lymphoma cells.

[0281] Experimental procedures: Four Raji ectopic mice were randomly selected, and two of them were directly given 100 μCi of 89 Zr-DFO-NYM1012 (non-blocking group), and two Raji ectopic mice were injected with unlabeled NYM1012 antibody through the tail vein, and the mass of the injected NYM1012 antibody was 89 Zr-DFO-NYM1012 is about 50 times the mass and was injected 3 hours after NYM1012 administration. 89 Zr-DFO-NYM1012 (100 μCi). Before scanning, mice were pre-anesthetized with an appropriate concentration of isoflurane / air mixture and placed in a MicroPET / CT imaging chamber (SNPC-303 Super Nova, Pingsheng Medical Technology (Kunshan) Co., Ltd.), and anesthesia was maintained with isoflurane / air mixture. 89 MicroPET / CT scans were performed 24, 48, and 96 hours after Zr-DFO-NYM1012 administration. Scanned images were reconstructed using the scanner software and analyzed using PMOD software.

[0282] Conclusion: Figure 4 As shown (the arrows indicate the tumor location), in the non-blocking group, 89 The radioactive uptake of Zr-DFO-NYM1012 was higher in Raji ectopic tumor tissues, whereas in the blocking group, 89 The radioactive uptake of Zr-DFO-NYM1012 was reduced in Raji ectopic tumor tissue, indicating that 89 Zr-DFO-NYM1012 has good tumor targeting properties.

[0283] Example 9: 89 Tissue distribution and targeting of Zr-DFO-NYM1012 in the Raji orthotopic mouse model

[0284] Mouse Model: The Raji orthotopic mouse model used in this experiment was provided by Biocytogen (Beijing) Pharmaceuticals Co., Ltd. This orthotopic model is based on the injection of B-luc-GFP Raji cells into B-NDG mice, a mouse tumor model established using human Burkitt's lymphoma cells. In this model, B-NDG mice are injected with Raji cells transduced with the luciferase gene and the GFP fluorescent group, and successful tumor engraftment is confirmed using fluorescence imaging.

[0285] Experimental procedures: 100 μCi of the above-mentioned 89 Zr-DFO-NYM1012. Before scanning, mice were pre-anesthetized with an appropriate concentration of isoflurane / air mixture and placed in a MicroPET / CT imaging chamber (SNPC-303Super Nova, Pingsheng Medical Technology (Kunshan) Co., Ltd.), and anesthesia was maintained with isoflurane / air mixture. 89 MicroPET / CT scans were performed 24, 48, and 96 hours after Zr-DFO-NYM1012 administration. Scanned images were reconstructed using the scanner software and analyzed using PMOD software.

[0286] Conclusion: Figure 5 As shown, 89 The biodistribution of Zr-DFO-NYM1012 in the Raji orthotopic model mice revealed high radioactivity uptake in tumors (joints and spine) and low uptake in other tissues, indicating that this radiolabeled antibody has good tumor targeting properties. 89 Zr-DFO-NYM1012 was able to detect lesions in an orthotopic model.

[0287] Example 10: Scientific research clinical trials

[0288] Patient information: Female, 77 years old, height: 1.55 cm, weight: 50 kg, diagnosed with multiple myeloma.

[0289] Institutional Ethics Committee Review: This study was reviewed and approved by the Ethics Committee of the Affiliated Hospital of Jiangnan University before it was performed.

[0290] Experimental procedure: 1.88 mCi of 89 Zr-DFO-NYM1012 was administered, and whole-body PET / CT static imaging was performed 24 and 72 hours after tracer administration.

[0291] Conclusion: Figure 6a and 6b As shown, 72 hours after injection, PET / CT imaging results showed multiple bone destruction throughout the body (including multiple sites in the skull, multiple sites in the spine, multiple sites in the bilateral ribs, multiple sites in the pelvis, sternum, bilateral scapulae, bilateral clavicles, bilateral humeri, and bilateral femurs) with abnormally increased CD38 expression. Combined with the patient's medical history, it is consistent with the manifestation of multiple myeloma. Abnormally increased radioactive uptake can be seen in the calcification of the spleen, and abnormally increased diffuse CD38 expression in the spleen, suggesting the possibility of reactive hyperplasia. It can be seen that 89 Zr-DFO-NYM1012 can detect human tumors and their micrometastases with clear imaging.

[0292] Example 11: Preparation of DTPA-NYM1012

[0293]

[0294] DTPA-NYM1012

[0295] p-SCN-Bn-CHX-A"-DTPA was purchased directly from Macrocyclics.

[0296] The antibody NYM1012 prepared in Example 2 was dissolved in a 0.1 mol / L NaHCO3 solution at a pH of 8.4 to a concentration of 10 mg / mL. The pH of the antibody solution was adjusted to 9.0 using a 0.1 mol / L Na2CO3 solution.

[0297] A 2 mmol / L DMSO solution of p-SCN-Bn-CHX-A"-DTPA ([(R)-2-amino-3-(4-isothiocyanatophenyl)propyl]-trans-(S,S)-cyclohexane-1,2-diamine-pentaacetic acid) was added to the above solution to achieve a molar ratio of p-SCN-Bn-CHX-A"-DTPA to antibody NYM1012 of approximately 5:1-10:1. After mixing, the mixture was reacted at 37°C for 60 minutes to obtain crude DTPA-NYM1012.

[0298] The crude DTPA-NYM1012 obtained above was purified using a PD10 column. After purification, 0.15 M acetic acid-sodium acetate solution (pH 7.2) was added to the collected fractions as a stabilizer, and the final solution was stored at -80°C for subsequent use.

[0299] The entire radiolabeling process was completed within 100 min, with a radiochemical yield of more than 80%.

[0300] Example 12: 177 Preparation of Lu-DTPA-NYM1012

[0301] The DTPA-NYM1012 synthesized above was used as chelate lutetium [ 177 Lu] precursor to obtain the precursor for SPECT / CT imaging by the following steps: 177 Lu-DTPA-NYM1012.

[0302] Add 10 μL of a solution containing approximately 10 mCi of radioactivity to 177 To the Lu hydrochloric acid solution, add 0.3 mL of acetic acid-sodium acetate buffer at a concentration of 0.15 mol / L and a pH of 7.2, and then add DTPA-NYM1012 (2 mg, in acetic acid-sodium acetate solution at pH 7.2). The mixture was mixed well and reacted at room temperature for 60 minutes (shaken every 10 minutes). The crude radiolabeled product was thus obtained. 177 Lu-DTPA-NYM1012.

[0303] The crude radiolabeled product obtained as described above was 177 Lu-DTPA-NYM1012 (if the total volume is less than 2.5 mL, the volume should be increased to 2.5 mL by adding the above-mentioned acetic acid-sodium acetate buffer solution) was added to a PD10 column that had been pre-equilibrated with 0.15 mol / L acetic acid-sodium acetate buffer (pH = 7.2) and all the equilibration effluent was discarded. Then, 3 mL of the above-mentioned acetic acid-sodium acetate buffer was added to collect all the effluent, which was filtered through a sterile 0.22 μm filter membrane to obtain the radioactive complex.177 Lu-DTPA-NYM1012.

[0304] Example 13: 177 Quality Control of Lu-DTPA-NYM1012

[0305] In this method, Radio-iTLC is used to control the radiolabeled product 177 The mass of Lu-DTPA-NYM1012 was determined using Radio-iTLC. 177 The radiochemical purity of Lu-DTPA-NYM1012 was determined using glass fiber paper as the TLC support and 0.5 M citrate / sodium citrate buffer (pH 5) as the developing solvent. 177 The radiochemical purity of Lu-DTPA-NYM1012 was higher than 99%.

[0306] The results were also confirmed by SEC-HPLC using 0.1 M PB + 0.2 M NaCl + 10% acetonitrile in water as the mobile phase. 177 The chemical and radiochemical purity of Lu-DTPA-NYM1012 was determined using a TSKgel G3000SWXL column with a flow rate of 0.8 mL / min and a detection wavelength of 280 nm. 177 The chemical purity of Lu-DTPA-NYM1012 was greater than 99%, and the radiochemical purity was found to be approximately 100%. Analytical results further revealed that the radiolabeled product maintained high chemical and radiochemical purity over 48 hours, with little or no release of free radionuclides, indicating good stability of the radiolabeled product.

[0307] Details of the radio-iTLC quality control method are as follows:

[0308] The TLC plate is essentially glass fiber paper, and the developing solvent is 0.5 M citric acid / sodium citrate buffer. The product sample is removed from the product container with a pipette gun and gently spotted on the surface of the glass fiber paper 1.5 cm from the bottom edge. The glass fiber paper is then placed in a test tube, in which citric acid buffer has been added for development. When the upward developing solvent front on the TLC reaches about 2.5 cm from the top of the chromatography paper, the TLC plate is removed from the test tube and dried, and analyzed using a Radio-TLC scanner. In this TLC system, free radionuclides ( 177 The Rf value of Lu) is between 0.8 and 1.0, while the Rf value of the radioactive product is between 0 and 0.2. The analysis results show that the radiolabeled product is of high purity and no free radionuclide is released within 48 hours, indicating good product stability.

[0309] Example 14: 177 Evaluation of the anti-tumor efficacy of Lu-DTPA-NYM1012 in MM.1S mouse model and RPMI8226 mouse model

[0310] Mouse Models: The RPMI8226 and MM.1S mouse models used in the experiments were provided by Hengjia Biotechnology (Suzhou) Co., Ltd. These are essentially ectopic tumor models based on subcutaneous injection of RPMI8226 and MM.1S tumor cells into BALB / c nude mice. These mouse models were established using human multiple myeloma peripheral blood B lymphocytes and human IgA-myeloma cells.

[0311] Experimental procedures: The RPMI8226 and MM.1S model mouse pools each consisted of 25 randomly selected model mice.

[0312] The tumor size of these mice was measured, and 40 mice with appropriate tumor volume were selected (20 RPMI8226 mouse models and 20 MM.1S mouse models). Each model group was further divided into two groups: 177 Lu treatment group and control group. In the RPMI8226 model study, 10 mice were selected to form 177 Lu treatment group, and the remaining 10 mice constituted the control group. In the MM.1S model study, 10 mice were selected to form 177 Lu treatment group, and the remaining 10 mice constituted the control group. 177 In the Lu treatment group, each animal was intravenously injected with a dose of approximately 400 μCi 177 Lu-DTPA-NYM1012 in saline solution. Each animal in the control group was injected intravenously with saline solution. The injection volume of the saline solution (control) group was close to 177 Lu treatment group. The injection time of each animal in each group was recorded.

[0313] exist 177 In the Lu treatment group and the control group, tumor volume (i.e., long and short diameters) and body weight were measured before and 2, 4, 6, 8, 10, 12, and 14 days after the administration of the treatment and control solutions, respectively. The physical condition of the mice was observed, and accurate records were made and kept.

[0314] Tumor volume was calculated using the long and short diameters of the tumors measured during the study period using the following formula:

[0315] Tumor volume (TV) = a × b 2 / 2 (where a is the major diameter and b is the minor diameter).

[0316] Conclusion: Figures 8a-8d As shown, compared with the control group,177 The tumor growth rate of RPMI8226 and MM.1S model mice in the Lu treatment group was slower, indicating that 177 Lu-DTPA-NYM1012 has good anti-tumor effect. 177 Treatment with Lu-DTPA-NYM1012 can more effectively inhibit tumor growth in RPMI8226 and MM.1S model mice.

[0317] Example 15: 177 Acute toxicity study of Lu-DTPA-NYM1012 in ICR mice

[0318] Mouse model: 6-8 week old female ICR mice were purchased from Hengjia Biotechnology (Suzhou) Co., Ltd.

[0319] Experimental procedures: Six mice were randomly selected and each ICR mouse was injected with 300-400 μCi of 177 Lu-DTPA-NYM1012. General animal health (hair condition, activity, and diet, etc.), mortality (time of death, etc.), and body weight were observed before dosing and 2, 4, 6, 8, 10, 12, and 14 days after dosing. On the final day of the experiment, all mice were dissected and their major organs were collected for observation and evaluation.

[0320] Conclusion: Figure 9 As shown, no death occurred and no abnormal reaction was observed during the 14-day observation period. Dissection of the collected organs showed no abnormalities, and the weight of all mice increased after the experiment compared to before the experiment. All of these indicate that 177 Lu-DTPA-NYM1012 has a good safety profile and meets the requirements for clinical studies for scientific research purposes.

[0321] The above embodiments are merely preferred embodiments for fully illustrating the present disclosure, and the scope of protection of the present disclosure is not limited thereto. Any equivalent substitution or transformation made by those skilled in the art based on the present disclosure is within the scope of protection of the present disclosure. The scope of protection of the present disclosure shall be subject to the claims.

[0322] Sequence Listing

[0323] SEQ ID NO: 1

[0324] RASSSVSSSA FSYVH

[0325] SEQ ID NO: 2

[0326] LASNLES

[0327] SEQ ID NO: 3

[0328] HHSRELPFT

[0329] SEQ ID NO: 4

[0330] LYWMN

[0331] SEQ ID NO: 5

[0332] KINPDSSTIN YTPSLKD

[0333] SEQ ID NO: 6

[0334] LWIATGGFDY

[0335] SEQ ID NO: 7

[0336] EIVMTQSPAS LSASLGQRAT ISCRASSSVS SSAFSYVHWY QQKSGQPPKL LIYLASNLESGVPARFSGSG SGTDFTLTIH PVESEDVATY YCHHSRELPF TFGSGTKLEI K

[0337] SEQ ID NO: 8

[0338] QVQLLESGGG LVQPGGSLKL SCVASGFDFS LYWMNWVRQA PGKGLEWIGK INPDSSTINYTPSLKDKFFI SRDNAKNTLY LQMTKVRSED TALYYCARLW IATGGFDYWG QGTTLTVSS

[0339] SEQ ID NO: 9

[0340] EIVMTQSPAS LSASLGQRAT ISCRASSSVS SSAFSYVHWY QQKSGQPPKL LIYLASNLESGVPARFSGSG SGTDFTLTIH PVESEDVATY YCHHSRELPF TFGSGTKLEI KRTVAAPSVF IFPPSDEQLKSGTASVVCLL NNFYPREAKV QWKVDNALQS GNSQESVTEQ DSKDSTYSLS STLTLSKADY EKHKVYACEVTHQGLSSPVT KSFNRGEC

[0341] SEQ ID NO: 10

[0342] QVQLLESGGG LVQPGGSLKL SCVASGFDFS LYWMNWVRQA PGKGLEWIGK INPDSSTINYTPSLKDKFFI SRDNAKNTLY LQMTKVRSED TALYYCARLW IATGGFDYWG QGTTLTVSSA STKGPSVFPLAPSSKSTSGG TAALGCLVKD YFPEPVTVSW NSGALTSGVH TFPAVLQSSG LYSLSSVVTV PSSSLGTQTYICNVNHKPSN TKVDKKVEPK SCDKTHTCPP CPAPELLGGP SVFLFPPKPK DTLMISRTPE VTCVVVDVSHEDPEVKFNWY VDGVEVHNAK TKPREEQYNS TYRVVSVLTV LHQDWLNGKE YKCKVSNKAL PAPIEKTISKAKGQPREPQV YTLPPSRDEL TKNQVSLTCL VKGFYPSDIA VEWESNGQPE NNYKTTPPVL DSDGSFFLYSKLTVDKSRWQ QGNVFSCSVM HEALHNHYTQ KSLSLSPGK

[0343] SEQ ID NO: 11

[0344] EIVMTQSPAS LSASLGQRAS ISCRASNSVS SSAYSYVHWY QQKSGQPPKL LIYLASNIQSGVPARFSGSG SGTDFTLTIH PVESEDVATY YCHHSRQLPS TFGSGTKLEI KRTVAAPSVF IFPPSDEQLKSGTASVVCLL NNFYPREAKV QWKVDNALQS GNSQESVTEQ DSKDSTYSLS STLTLSKADY EKHKVYACEVTHQGLSSPVT KSFNRGEC

[0345] SEQ ID NO: 12

[0346] QVQLLESGGG LVQPGGSLKL SCVASGYNFS LYYMNWVRQA PGKGLEWIGK IQPESSTIQYTPSLKDKFFI SRDNAKNTLY LQMTKVRSED TALYYCARLW IGSGGFDYWG QGTTLTVSSA STKGPSVFPLAPSSKSTSGG TAALGCLVKD YFPEPVTVSW NSGALTSGVH TFPAVLQSSG LYSLSSVVTV PSSSLGTQTYICNVNHKPSN TKVDKKVEPK SCDKTHTCPP CPAPELLGGP SVFLFPPKPK DTLMISRTPE VTCVVVDVSHEDPEVKFNWY VDGVEVHNAK TKPREEQYNS TYRVVSVLTV LHQDWLNGKE YKCKVSNKAL PAPIEKTISKAKGQPREPQV YTLPPSRDEL TKNQVSLTCL VKGFYPSDIA VEWESNGQPE NNYKTTPPVL DSDGSFFLYSKLTVDKSRWQ QGNVFSCSVM HEALHNHYTQ KSLSLSPGK

[0347] SEQ ID NO: 13

[0348] EIVMTQSPAS LSASLGQRAT ISCRASSSVS SSAYSYVHWY QQKSGQPPKL LIYLASDLQSGVPARFSGSG SGTDFTLTIH PVESEDVATY YCHHSRELPY SFGSGTKLEI KRTVAAPSVF IFPPSDEQLKSGTASVVCLL NNFYPREAKV QWKVDNALQS GNSQESVTEQ DSKDSTYSLS STLTLSKADY EKHKVYACEVTHQGLSSPVT KSFNRGEC

[0349] SEQ ID NO: 14

[0350] QVQLLESGGG LVQPGGSLKL SCVASGFDFS LYWMNWVRQA PGKGLEWIGK ISPNSSTINYSPSLKDKFFI SRDNAKNTLY LQMTKVRSED TALYYCARLY IASGGFDYWG QGTTLTVSSA STKGPSVFPLAPSSKSTSGG TAALGCLVKD YFPEPVTVSW NSGALTSGVH TFPAVLQSSG LYSLSSVVTV PSSSLGTQTYICNVNHKPSN TKVDKKVEPK SCDKTHTCPP CPAPELLGGP SVFLFPPKPK DTLMISRTPE VTCVVVDVSHEDPEVKFNWY VDGVEVHNAK TKPREEQYNS TYRVVSVLTV LHQDWLNGKE YKCKVSNKAL PAPIEKTISKAKGQPREPQV YTLPPSRDEL TKNQVSLTCL VKGFYPSDIA VEWESNGQPE NNYKTTPPVL DSDGSFFLYSKLTVDKSRWQ QGNVFSCSVM HEALHNHYTQ KSLSLSPGK

[0351] SEQ ID NO: 15

[0352] EIVMTQSPAS LSASLGQRAT ISCRASNSVS TSAFSYVHWY QQKSGQPPKL LIYLASNLESGVPARFSGSG SGTDFTLTIH PVESEDVATY YCHHSREVPF TFGSGTKLEI KRTVAAPSVF IFPPSDEQLKSGTASVVCLL NNFYPREAKV QWKVDNALQS GNSQESVTEQ DSKDSTYSLS STLTLSKADY EKHKVYACEVTHQGLSSPVT KSFNRGEC

[0353] SEQ ID NO: 16

[0354] QVQLLESGGG LVQPGGSLKL SCVASGFDFS LYWMNWVRQA PGKGLEWIGK ISPDSSSLNYTPSVKDKFFI SRDNAKNTLY LQMTKVRSED TALYYCARLW IATGGYNYWG QGTTLTVSSA STKGPSVFPLAPSSKSTSGG TAALGCLVKD YFPEPVTVSW NSGALTSGVH TFPAVLQSSG LYSLSSVVTV PSSSLGTQTYICNVNHKPSN TKVDKKVEPK SCDKTHTCPP CPAPELLGGP SVFLFPPKPK DTLMISRTPE VTCVVVDVSHEDPEVKFNWY VDGVEVHNAK TKPREEQYNS TYRVVSVLTV LHQDWLNGKE YKCKVSNKAL PAPIEKTISKAKGQPREPQV YTLPPSRDEL TKNQVSLTCL VKGFYPSDIA VEWESNGQPE NNYKTTPPVL DSDGSFFLYSKLTVDKSRWQ QGNVFSCSVM HEALHNHYTQ KSLSLSPGK

[0355] SEQ ID NO: 17

[0356] EIVMTQSPAS LSASLGQRAX ISCRASXSVS XSAXSYVHWY QQKSGQPPKL LIYLASXXXSGVPARFSGSG SGTDFTLTIH PVESEDVATY YCHHSRXXPX XFGSGTKLEI KRTVAAPSVF IFPPSDEQLKSGTASVVCLL NNFYPREAKV QWKVDNALQS GNSQESVTEQ DSKDSTYSLS STLTLSKADY EKHKVYACEVTHQGLSSPVT KSFNRGEC

[0357] SEQ ID NO: 18

[0358] QVQLLESGGG LVQPGGSLKL SCVASGXXFS LYXMNWVRQA PGKGLEWIGK IXPXSSXXXYXPSXKDKFFI SRDNAKNTLY LQMTKVRSED TALYYCARLX IXXGGXXYWG QGTTLTVSSA STKGPSVFPLAPSSKSTSGG TAALGCLVKD YFPEPVTVSW NSGALTSGVH TFPAVLQSSG LYSLSSVVTV PSSSLGTQTYICNVNHKPSN TKVDKKVEPK SCDKTHTCPP CPAPELLGGP SVFLFPPKPK DTLMISRTPE VTCVVVDVSHEDPEVKFNWY VDGVEVHNAK TKPREEQYNS TYRVVSVLTV LHQDWLNGKE YKCKVSNKAL PAPIEKTISKAKGQPREPQV YTLPPSRDEL TKNQVSLTCL VKGFYPSDIA VEWESNGQPE NNYKTTPPVL DSDGSFFLYSKLTVDKSRWQ QGNVFSCSVM HEALHNHYTQ KSLSLSPGK

[0359] SEQ ID NO: 19

[0360] gagatcgtga tgacccagag ccctgccagc ctgagcgcca gcctgggcca gagggccaccatcagctgca gggccagcag cagcgtgagc agcagcgcct tcagctacgt gcactggtac cagcagaagagcggccagcc tcctaagctg ctgatctacc tggccagcaa cctggagagc ggcgtgcctg ccaggttcagcggcagcggc agcggcaccg acttcaccct gaccatccac cctgtggaga gcgaggacgt ggccacctactactgccacc acagcaggga gctgcctttc accttcggca gcggcaccaa gctggagatc aag

[0361] SEQ ID NO: 20

[0362] caggtgcagc tgctggagag cggcggcggc ctggtgcagc ctggcggcag cctgaagctgagctgcgtgg ccagcggctt cgacttcagc ctgtactgga tgaactgggt gaggcaggcc cctggcaagggcctggagtg gatcggcaag atcaaccctg acagcagcac catcaactac acccctagcc tgaaggacaagttcttcatc agcagggaca acgccaagaa caccctgtac ctgcagatga ccaaggtgag gagcgaggacaccgccctgt actactgcgc caggctgtgg atcgccaccg gcggcttcga ctactggggc cagggcaccaccctgaccgt gagcagc

[0363] SEQ ID NO: 21

[0364] gagatcgtga tgacccagag ccctgccagc ctgagcgcca gcctgggcca gagggccaccatcagctgca gggccagcag cagcgtgagc agcagcgcct tcagctacgt gcactggtac cagcagaagagcggccagcc tcctaagctg ctgatctacc tggccagcaa cctggagagc ggcgtgcctg ccaggttcagcggcagcggc agcggcaccg acttcaccct gaccatccac cctgtggaga gcgaggacgt ggccacctactactgccacc acagcaggga gctgcctttc accttcggca gcggcaccaa gctggagatc aagcgtacggtggctgcacc atctgtcttc atcttcccgc catctgatga gcagttgaaa tctggaactg cctctgttgtgtgcctgctg aataacttct atcccagaga ggccaaagta cagtggaagg tggataacgc cctccaatcgggtaactccc aggagagtgt cacagagcag gacagcaagg acagcaccta cagcctcagc agcaccctgacgctgagcaa agcagactac gagaaacaca aagtctacgc ctgcgaagtc acccatcagg gcctgagctcgcccgtcaca aagagcttca acaggggaga gtgttag

[0365] SEQ ID NO: 22

[0366] caggtgcagc tgctggagag cggcggcggc ctggtgcagc ctggcggcag cctgaagctgagctgcgtgg ccagcggctt cgacttcagc ctgtactgga tgaactgggt gaggcaggcc cctggcaagggcctggagtg gatcggcaag atcaaccctg acagcagcac catcaactac acccctagcc tgaaggacaagttcttcatc agcagggaca acgccaagaa caccctgtac ctgcagatga ccaaggtgag gagcgaggacaccgccctgt actactgcgc caggctgtgg atcgccaccg gcggcttcga ctactggggc cagggcaccaccctgaccgt gagcagcgct agcaccaagg gcccatcggt cttccccctg gcaccctcct ccaagagcacctctgggggc acagcggccc tgggctgcct ggtcaaggac tacttccccg aaccggtgac ggtgtcgtggaactcaggcg ccctgaccag cggcgtgcac accttcccgg ctgtcctaca gtcctcagga ctctactccctcagcagcgt ggtgaccgtg ccctccagca gcttgggcac ccagacctac atctgcaacg tgaatcacaagcccagcaac accaaggtgg acaagaaagt tgagcccaaa tcttgtgaca aaactcacac atgcccaccgtgcccagcac ctgaactcct ggggggaccg tcagtcttcc tcttcccccc aaaacccaag gacaccctcatgatctcccg gacccctgag gtcacgtgcg tggtggtgga cgtgagccac gaagaccccg aggtcaagttcaactggtac gtggacggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gtacaacagcacgtaccgtg tggtcagcgtcctcaccgtc ctgcaccagg actggctgaa tggcaaggag tacaagtgca aggtctccaa caaagccctc ccagccccca tcgagaaaac catctccaaa gccaaagggc agccccgaga accacaggtg tacaccctgc ccccatcccg ggatgagctg accaagaacc aggtcagcct gacctgcctggtcaaaggct tctatcccag cgacatcgcc gtggagtggg agagcaatgg gcagccggag aacaactacaagaccacgcc tcccgtgctg gactccgacg gctccttctt cctctacagc aagctcaccg tggacaagagcaggtggcag caggggaacg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacgcagaagagcctct ccctgtctcc gggtaaatga

Claims

1. A radiolabeled antibody conjugate represented by Formula I: Ab-(L-M) p (I) in, Ab is an antibody or antigen-binding fragment that specifically binds to CD38 protein. L is the connector, M is the radionuclide, and p is an integer of 1 to 10, preferably 2 to 8, preferably 4 to 8, preferably 3 to 5.

2. The radiolabeled antibody conjugate according to claim 1, wherein L is represented by formula II Lk-YT (II) in, Lk does not exist or is a coupling group, such as -C(=S)NH-, -C(=O)NH-, -C(=S)-, -C(=O)-, -S-, -CH2C(=O)-, or , Y is a spacer, and T is a chelating group.

3. The radiolabeled antibody conjugate according to claim 2, which is represented by one of the following structures: 、 、 、 、 、 、 or in, Y is independently selected from a bond, a C1-C 20 Alkylene, C2-C 20 Alkenylene, C2-C 20 Alkynylidene, C1-C 20 Heteroalkylene, C3-C 20 Cycloalkylene, C3-C 20 Heterocyclylene, C6-C 20 Arylene or C5-C 20 heteroarylene, keto, amino, thio, -(C=S)-, -O-, or a combination thereof, optionally substituted with one or more halogen, cyano, hydroxy, alkoxy, monocyclic cycloalkyl, monocyclic heterocyclyl, T is independently selected from a ligand derived from any of: 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA), S-2-methyl-1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid, N,N''-bis[2-hydroxy-5-(carboxyethyl)benzyl]ethylenediamine-N,N''-diacetic acid (HBED-CC), 1,4,7-Triazacyclononane-1,4,7-triacetic acid (NOTA), 2-(4,7-bis(carboxymethyl)-1,4,7-triazanonan-1-yl)glutaric acid (NODAGA), 2-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododec-1-yl)pentanedioic acid (DOTAGA), 1,4,7-Triazacyclononanephosphinic acid (TRAP), 1,4,7-Triazacyclononane-1-[methylene(2-carboxyethyl)phosphinic acid]-4,7-bis[methylene(2-hydroxymethyl)phosphinic acid](NOPO), 3,6,9,15-tetraazabicyclo[9.3.1.]pentadeca-1(15),11,13-triene-3,6,9-triacetic acid (PCTA), N'-{5-[acetyl(hydroxy)amino]pentyl}-N-[5-({4-[(5-aminopentyl)(hydroxy)amino]-4-oxobutanoyl}amino)pentyl]-N-hydroxysuccinamide (DFO), Diethylenetriaminepentaacetic acid (DTPA), trans-cyclohexyl-diethylenetriaminepentaacetic acid (CHX-DTPA), 1-oxa-4,7,10-triazacyclododecane-4,7,10-triacetic acid (oxo-Do3A), N x S 4-x (such as N4, N2S2, N3S), or a combination thereof, M is selected from diagnostic radionuclides, therapeutic radionuclides and non-radioactive elements, Preferably, the diagnostic radionuclides are independently selected from 68 Ga, 99m Tc, 89 Zr, 43 Sc, 111 In, 45 Ti, 52 Mn, 59 Fe, 64 Cu, 94m Tc, 67 Ga, 71 As、 72 As、 74 As、 82m Rb, 86 Y or a combination thereof, Preferably, the therapeutic radionuclides are independently selected from 177 Lu, 90 Y. 153 Sm, 67 Cu, 89 Sr. 166 Ho, 177 Yb, 47 Sc, 186 Re、 188 Re、 212 Bi, 213 Bi, 149 Pm, 212 Pb, 211 At 223 Ra, 225 Ac, 227 Th, 161 Tb or a combination thereof, Preferably, the non-radioactive elements are independently selected from Ga, Fe or a combination thereof, Independently selected from covalent bonds, ionic bonds, van der Waals forces, conjugated bonds or combinations thereof.

4. The radiolabeled antibody conjugate according to claim 3, which is represented by one of the following structures: 、 or , preferably .

5. The radiolabeled antibody conjugate according to claim 3 or 4, in, Y is independently selected from: 、 、 、 、 、 、 、 、 、 、 、 、 、 or a combination thereof, wherein R is independently selected from hydrogen or C1-C 10 Alkyl, n is an integer from 0 to 10.

6. The radiolabeled antibody conjugate according to any one of claims 3 to 5, in, Y is independently selected from a bond, 、 、 、 、 or a combination thereof.

7. The radiolabeled antibody conjugate of any one of claims 3 to 6, wherein T is independently selected from DFO, DOTA, DTPA, CHX-DTPA, p-SCN-Bn-DFO, p-NH2-Bn-CHX-A"-DTPA, p-SCN-Bn-CHX-A"-DTPA, or a combination thereof.

8. The radiolabeled antibody conjugate according to any one of claims 3 to 7, wherein Lk-YT is independently selected from any one of the following: 1-(4-isothiocyanatophenyl)-3-[6,17-dihydroxy-7,10,18,21-tetraoxy-27-(N-acetylhydroxyamino)-6,11,17,22-tetraazaheptadecanoyl]thiourea (p-SCN-Bn-DFO), [(R)-2-amino-3-(4-aminophenyl)propyl]-trans-(S,S)-cyclohexane-1,2-diamine-pentaacetic acid (p-NH2-Bn-CHX-A''-DTPA), [(R)-2-amino-3-(4-isothiocyanatophenyl)propyl]-trans-(S,S)-cyclohexane-1,2-diamine-pentaacetic acid (p-SCN-Bn-CHX-A''-DTPA), 1-(p-isothiocyanatobenzyl)-3-methyl-DTPA (1B3M), Benzyl isothiocyanate-DTPA (SCN-Bz-DTPA), 1-(2)-methyl-4-isocyanatobenzyl-DTPA (MX-DTPA), Mercaptoacetyl triglycine (MAG3), Mercaptoacetyl diglycine (MAG2), DOTAGA, NOTA, NODAGA, DOTA and HBED-CC, Preferably, Lk-YT is independently selected from any one of the following: DOTAGA, NOTE、 NODAGA DOTA, HBED-CC, p-SCN-Bn-DFO, p-NH2-Bn-CHX-A"-DTPA, p-SCN-Bn-CHX-A"-DTPA, Preferably, Lk-YT is independently selected from any one of the following: p-SCN-Bn-DFO, p-NH2-Bn-CHX-A"-DTPA, p-SCN-Bn-CHX-A"-DTPA or a combination thereof.

9. The radiolabeled antibody conjugate according to any one of claims 3 to 8, wherein M is independently selected from 99m Tc, 89 Zr, 177 Lu, 225 Ac or a combination thereof.

10. The radiolabeled antibody conjugate according to any one of claims 1 to 9, which is represented by any one of the following structures: 、 、 、 、 、 or .

11. The radiolabeled antibody conjugate according to any one of claims 1 to 10, wherein Ab is selected from the group consisting of a monoclonal antibody, a single-chain antibody, a chimeric antibody, a humanized antibody, and a fully human antibody.

12. The radiolabeled antibody conjugate of any one of claims 1 to 11, wherein the antigen-binding fragment is selected from the group consisting of Fab, Fab', F(ab)2, F(ab')2, Fv, and ScFv fragments.

13. The radiolabeled antibody conjugate according to any one of claims 1 to 12, wherein Ab comprises an antibody light chain or a fragment thereof, wherein the antibody light chain or fragment thereof comprises LCDR1-3, wherein the LCDR1-3 comprises LCDR1, LCDR2, and LCDR3, wherein the LCDR1 comprises the amino acid sequence as shown in SEQ ID NO: 1; the LCDR2 comprises the amino acid sequence as shown in SEQ ID NO: 2; and the LCDR3 comprises the amino acid sequence as shown in SEQ ID NO:

3.

14. The radiolabeled antibody conjugate according to claim 13, wherein the light chain of the antibody or a fragment thereof comprises a light chain variable region VL, and the light chain variable region VL comprises the amino acid sequence shown in SEQ ID NO:

7.

15. The radiolabeled antibody conjugate of claim 13, wherein the light chain of the antibody or a fragment thereof comprises the amino acid sequence shown in SEQ ID NO:

17.

16. The radiolabeled antibody conjugate of claim 14, wherein the light chain of the antibody or a fragment thereof comprises the amino acid sequence shown in any one of SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13 and SEQ ID NO:

15.

17. The radiolabeled antibody conjugate of any one of claims 1 to 16, wherein Ab comprises a heavy chain of an antibody or a fragment thereof, the heavy chain of the antibody or the fragment thereof comprising HCDR1, HCDR2, and HCDR3, the HCDR1 comprising the amino acid sequence as shown in SEQ ID NO: 4; the HCDR2 comprising the amino acid sequence as shown in SEQ ID NO: 5; and the HCDR3 comprising the amino acid sequence as shown in SEQ ID NO:

6.

18. The radiolabeled antibody conjugate of claim 17, wherein the heavy chain of the antibody or a fragment thereof comprises a heavy chain variable region VH, and the heavy chain variable region VH comprises the amino acid sequence shown in SEQ ID NO:

8.

19. The radiolabeled antibody conjugate of claim 17, wherein the heavy chain of the antibody comprises the amino acid sequence shown in SEQ ID NO:

18.

20. The radiolabeled antibody conjugate of claim 19, wherein the heavy chain of the antibody comprises the amino acid sequence shown in any one of SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14 and SEQ ID NO:

16.

21. A pharmaceutical composition comprising the radiolabeled antibody conjugate according to any one of claims 1 to 20 and a pharmaceutically acceptable carrier.

22. A kit comprising a radionuclide M and a precursor Ab-(Lk-Y-T') p wherein said M, Ab, Lk, Y and p have the same meanings as defined in any one of claims 1-20, and T' is independently selected from any one of the following: 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA), S-2-methyl-1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid, N,N''-bis[2-hydroxy-5-(carboxyethyl)benzyl]ethylenediamine-N,N''-diacetic acid (HBED-CC), 1,4,7-Triazacyclononane-1,4,7-triacetic acid (NOTA), 2-(4,7-bis(carboxymethyl)-1,4,7-triazanonan-1-yl)glutaric acid (NODAGA), 2-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododec-1-yl)pentanedioic acid (DOTAGA), 1,4,7-Triazacyclononanephosphinic acid (TRAP), 1,4,7-Triazacyclononane-1-[methylene(2-carboxyethyl)phosphinic acid]-4,7-bis[methylene(2-hydroxymethyl)phosphinic acid](NOPO), 3,6,9,15-tetraazabicyclo[9.3.1.]pentadeca-1(15),11,13-triene-3,6,9-triacetic acid (PCTA), N'-{5-[acetyl(hydroxy)amino]pentyl}-N-[5-({4-[(5-aminopentyl)(hydroxy)amino]-4-oxobutanoyl}amino)pentyl]-N-hydroxysuccinamide (DFO), Diethylenetriaminepentaacetic acid (DTPA), trans-cyclohexyl-diethylenetriaminepentaacetic acid (CHX-DTPA), p-Oxa-4,7,10-triazacyclododecane-4,7,10-triacetic acid (oxo-Do3A), N x S 4-x (such as N4, N2S2, N3S), or a combination thereof; Preferably, Lk-YT is independently selected from any one of the following: 1-(4-isothiocyanatophenyl)-3-[6,17-dihydroxy-7,10,18,21-tetraoxy-27-(N-acetylhydroxyamino)-6,11,17,22-tetraazaheptadecanoyl]thiourea (p-SCN-Bn-DFO), [(R)-2-amino-3-(4-aminophenyl)propyl]-trans-(S,S)-cyclohexane-1,2-diamine-pentaacetic acid (p-NH2-Bn-CHX-A''-DTPA), [(R)-2-amino-3-(4-isothiocyanatophenyl)propyl]-trans-(S,S)-cyclohexane-1,2-diamine-pentaacetic acid (p-SCN-Bn-CHX-A''-DTPA), 1-(p-isothiocyanatobenzyl)-3-methyl-DTPA (1B3M), Benzyl isothiocyanate-DTPA (SCN-Bz-DTPA), 1-(2)-methyl-4-isocyanatobenzyl-DTPA (MX-DTPA), Mercaptoacetyl triglycine (MAG3), Mercaptoacetyl diglycine (MAG2), DOTAGA, NOTA, NODAGA, DOTA and HBED-CC, Preferably, Lk-YT is independently selected from any one of the following: p-SCN-Bn-DFO, p-NH2-Bn-CHX-A"-DTPA, p-SCN-Bn-CHX-A"-DTPA or a combination thereof.

23. A method for preparing a radiolabeled antibody conjugate according to any one of claims 1 to 20, comprising reacting the precursor Ab-(Lk-Y-T') p Reacts with radionuclide M.

24. A method having Ab-(Lk-Y-T') p An antibody conjugate having a structure of wherein Ab, Lk, Y and p have the same meanings as defined in any one of claims 1 to 20, and T' is independently selected from any one of the following: 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA), S-2-methyl-1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid, N,N''-bis[2-hydroxy-5-(carboxyethyl)benzyl]ethylenediamine-N,N''-diacetic acid (HBED-CC), 1,4,7-Triazacyclononane-1,4,7-triacetic acid (NOTA), 2-(4,7-bis(carboxymethyl)-1,4,7-triazanonan-1-yl)glutaric acid (NODAGA), 2-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododec-1-yl)pentanedioic acid (DOTAGA), 1,4,7-Triazacyclononanephosphinic acid (TRAP), 1,4,7-Triazacyclononane-1-[methylene(2-carboxyethyl)phosphinic acid]-4,7-bis[methylene(2-hydroxymethyl)phosphinic acid](NOPO), 3,6,9,15-tetraazabicyclo[9.3.1.]pentadeca-1(15),11,13-triene-3,6,9-triacetic acid (PCTA), N'-{5-[acetyl(hydroxy)amino]pentyl}-N-[5-({4-[(5-aminopentyl)(hydroxy)amino]-4-oxobutanoyl}amino)pentyl]-N-hydroxysuccinamide (DFO), Diethylenetriaminepentaacetic acid (DTPA), trans-cyclohexyl-diethylenetriaminepentaacetic acid (CHX-DTPA), p-Oxa-4,7,10-triazacyclododecane-4,7,10-triacetic acid (oxo-Do3A), N x S 4-x (such as N4, N2S2, N3S), or a combination thereof; Preferably, Lk-YT is independently selected from any one of the following: 1-(4-isothiocyanatophenyl)-3-[6,17-dihydroxy-7,10,18,21-tetraoxy-27-(N-acetylhydroxyamino)-6,11,17,22-tetraazaheptadecanoyl]thiourea (p-SCN-Bn-DFO), [(R)-2-amino-3-(4-aminophenyl)propyl]-trans-(S,S)-cyclohexane-1,2-diamine-pentaacetic acid (p-NH2-Bn-CHX-A''-DTPA), [(R)-2-amino-3-(4-isothiocyanatophenyl)propyl]-trans-(S,S)-cyclohexane-1,2-diamine-pentaacetic acid (p-SCN-Bn-CHX-A''-DTPA), 1-(p-isothiocyanatobenzyl)-3-methyl-DTPA (1B3M), Benzyl isothiocyanate-DTPA (SCN-Bz-DTPA), 1-(2)-methyl-4-isocyanatobenzyl-DTPA (MX-DTPA), Mercaptoacetyl triglycine (MAG3), Mercaptoacetyl diglycine (MAG2), DOTAGA, NOTA, NODAGA, DOTA and HBED-CC, Preferably, Lk-YT is independently selected from any one of the following: p-SCN-Bn-DFO, p-NH2-Bn-CHX-A"-DTPA, p-SCN-Bn-CHX-A"-DTPA or a combination thereof.

25. A pharmaceutical composition comprising the radiolabeled antibody conjugate of any one of claims 1 to 20 and the antibody conjugate of claim 24.

26. Use of the radiolabeled antibody conjugate according to any one of claims 1 to 20, the pharmaceutical composition according to claim 21 or 25, or the kit according to claim 22 in the preparation of a medicament for detecting or assisting in the detection of CD38.

27. A method of imaging, comprising: administering to an animal the radiolabeled antibody conjugate of any one of claims 1 to 20 or the pharmaceutical composition of claim 21 or 25; and detecting the presence of the radiolabeled antibody conjugate in vivo by imaging.

28. The method of claim 27, wherein the presence of the radiolabeled antibody conjugate is detected by positron emission tomography (PET) or single photon emission computed tomography (SPECT).

29. A method of treating cancer comprising administering to a subject the radiolabeled antibody conjugate of any one of claims 1 to 20 or the pharmaceutical composition of claim 21 or 25.

30. The method of claim 29, wherein the subject is a CD38 positive subject.

31. The method of claim 29 or 30, wherein the cancer is a solid tumor.

32. The method of claim 31 , wherein the cancer is selected from the group consisting of brain cancer, renal cell carcinoma, ovarian cancer, bladder cancer, prostate cancer, breast cancer, hepatocellular carcinoma, bone cancer, colon cancer, non-small cell lung cancer, head and neck squamous cell carcinoma, colorectal cancer, mesothelioma, B-cell lymphoma, and melanoma.

33. The method of claim 29, wherein the cancer is a hematological cancer.

34. The method of claim 33, wherein the cancer is selected from the group consisting of myeloma, lymphoma, and leukemia, preferably, the cancer is selected from the group consisting of acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), chronic myelomonocytic leukemia (CMML), Hodgkin's lymphoma, non-Hodgkin's lymphoma, and multiple myeloma.

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