CN114437215B - Anti-human CD38 antibody and preparation method and application thereof - Google Patents

Abstract

The invention discloses an antibody capable of binding to human CD38, and a preparation method and application thereof. The antibody of the invention has anti-human CD38 antibodies with different affinities of high, medium and low, can be combined with CD38 of cynomolgus monkey, and can kill tumor cells through an Fc-dependent immune effector mechanism. Can be used for treating CD38 related diseases, such as multiple myeloma, non-Hodgkin's lymphoma, AL type amyloidosis, T cell lymphoma, and diffuse large B cell lymphoma.

Description

Anti-human CD38 antibody and its preparation method and use

technical field

The present invention relates to the field of antibodies, more specifically, the present invention relates to an anti-human CD38 antibody and its preparation method and application.

Background technique

CD38 is a 46kDa type II transmembrane glycoprotein with an extracellular segment consisting of 258 amino acids. The functions of CD38 include receptor-mediated adhesion and signal transduction and bifunctional in vitro enzyme activity to promote intracellular calcium mobilization. Under normal conditions, CD38 is expressed at low levels on lymphoid and myeloid cells, but in multiple myeloma, malignant tumor cells specifically express CD38, making CD38 an ideal myeloma target.

Multiple myeloma is a malignant plasma cell disease that occurs in B lymphocytes. Usually, myeloma cells proliferate clonally in the bone marrow and bone spongy soft tissue, causing osteolytic bone destruction. The poor prognosis is often accompanied by anemia, renal failure, and various damages caused by extramedullary infiltration of myeloma cells. The incidence of multiple myeloma in China has surpassed that of acute leukemia, and it is the second hematological malignancy after non-Hodgkin's lymphoma. There are about 15,000 to 20,000 new cases of MM in China every year, the median survival period of patients is about 4 to 5 years, and the number of patients in China is about 80,000 to 100,000. With the arrival of an aging society and the deterioration of the environment and other factors, the incidence of multiple myeloma has been found to increase.

Studies have found that CD38 monoclonal antibodies mainly kill tumor cells through Fc-dependent immune effector mechanisms, including complement-mediated cytotoxicity (CDC), antibody-mediated cytotoxicity (ADCC), antibody-mediated phagocytosis ( ADCP), and through apoptosis. In addition, CD38 antibody has an immunomodulatory effect: it regulates myeloma cells by reducing CD38+ immunosuppression, regulating cells, and promoting T cell expansion and activity.

Internationally, anti-CD38 antibody drugs such as Johnson & Johnson's Daratumumab and Sanofi's Isatuximab are already on the market, and I-Mab's MOR202 is in clinical trials. Antibodies targeting CD38 have very significant curative effect advantages in the treatment of multiple myeloma compared with current drugs, macromolecule and small molecule drugs under research. Good competitive advantage.

At present, the main treatment methods for myeloma in my country are stem cell transplantation and Janssen's Velcade (bortezomib) or Celgene's REVLIMIDE (lenalidomide) and other combination therapy. In the past two years, the FDA has approved a number of new drug molecules for the treatment of multiple myeloma. With the continuous emergence of new drugs for treatment, the curative effect has been improved and the prognosis has been improved. The current median survival period is 7-8 years, and the multiple myeloma is gradually becoming The transformation of myeloma into a chronic disease objectively drives the growth of the multiple myeloma treatment market. The arrival of CD38 monoclonal antibody provides a new option for multiple myeloma patients, especially relapsed and refractory patients. We look forward to more breakthroughs in the field of CD38 antibody research to benefit more patients.

Contents of the invention

The inventors of the present invention have conducted a large number of experiments and obtained a group of monoclonal antibodies that can specifically bind to CD38 on the cell surface, and these antibodies can also bind to CD38 in cynomolgus monkeys. A series of anti-human CD38 antibodies with high, medium and low affinity were obtained, which are able to kill tumor cells through Fc-dependent immune effector mechanisms.

In a first aspect, the present application provides an antibody or an antigen-binding portion thereof that specifically binds to human CD38, comprising a heavy chain variable region, the heavy chain variable region comprising HCDR3 sequence, optionally further comprising HCDR1 and/or or HCDR2 sequence. In some embodiments, the aforementioned HCDR3 sequence comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, 39, 42, 45 and 48 . In some embodiments, the aforementioned HCDR2 sequence comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 5, 8, 11, 14, 17, 20, 23, 26, 29, 32, 35, 38, 41, 44 and 47 . In some embodiments, the aforementioned HCDR1 sequence comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40, 43 and 46 .

In some embodiments, the heavy chain variable region described above comprises an amino acid sequence having at least 80% homology to an amino acid sequence selected from the group consisting of SEQ ID Nos: 97, 99, 101, 103, 105, 107, 109 and 111, or The heavy chain variable region comprises an amino acid sequence selected from SEQ ID NOs: 97, 99, 101, 103, 105, 107, 109 and 111.

In some embodiments, the antibody, or antigen-binding portion thereof, that specifically binds human CD38 further comprises a light chain variable region, wherein the light chain variable region comprises LCDR1, LCDR2 and/or LCDR3 sequences. In certain embodiments, the LCDR1 sequence comprises a sequence selected from SEQ ID NOs: 49, 52, 55, 58, 61, 64, 67, 70, 73, 76, 79, 82, 85, 88, 91 and 94 amino acid sequence. In some embodiments, the LCDR1 sequence further comprises an amino acid sequence selected from SEQ ID NO:115. In certain embodiments, the LCDR2 sequence comprises a sequence selected from SEQ ID NOs: 50, 53, 56, 59, 62, 65, 68, 71, 74, 77, 80, 83, 86, 89, 92 and 95 amino acid sequence. In some embodiments, the LCDR2 sequence further comprises an amino acid sequence selected from SEQ ID NO:116. In certain embodiments, the LCDR3 sequence comprises a sequence selected from SEQ ID NOs: 51, 54, 57, 60, 63, 66, 69, 72, 75, 78, 81, 84, 87, 90, 93 and 96 amino acid sequence. In some embodiments, the LCDR3 sequence further comprises an amino acid sequence selected from SEQ ID NO:117.

In some embodiments, the light chain variable region described above comprises an amino acid sequence having at least 80% homology to an amino acid sequence selected from the group consisting of SEQ ID NOs: 98, 100, 102, 104, 106, 108, 110 and 112; or The light chain variable region comprises an amino acid sequence selected from SEQ ID NOs:98, 100, 102, 104, 106, 108, 110 and 112.

In an alternative embodiment, the above-mentioned antigen binding portion is selected from Fab fragments, Fab' fragments, F(ab')2 fragments, Fv fragments, scFv fragments, Fd fragments and single domain antibodies.

In some embodiments, the antibody specifically binding to human CD38 described in the first aspect is a murine monoclonal antibody.

In some embodiments, the antibody specifically binding to human CD38 described in the first aspect is a humanized antibody.

In the second aspect, the present application provides an expression vector, and the nucleotide molecule contained in the expression vector can encode the above-mentioned amino acid sequence.

In some embodiments, the expression vector is pTT5, pUC57, pDR1, pcDNA3.1(+), pDHFF or pCHO 1.0, etc.

In a third aspect, the present application provides a host cell containing the above-mentioned expression vector. In some embodiments, the host cell is HEK293, COS, CHO, NSO, sf9, sf21, DH5α, BL21(DE3) or TG1, etc.

In a fourth aspect, the present application provides a method for preparing the antibody or antigen-binding portion thereof that specifically binds to human CD38 described in the first aspect, the method comprising the following steps:

a) culturing the host cell of the third aspect under expression conditions that enable the host cell to produce the antibody or antigen-binding portion thereof, thereby expressing the antibody or antigen-binding portion thereof; and

b) isolating and purifying said antibody or antigen-binding portion thereof expressed in a).

In a fifth aspect, the present application provides a pharmaceutical composition, which comprises the anti-human CD38 antibody or antigen-binding portion thereof described in the first aspect and a pharmaceutically acceptable carrier.

In some embodiments, the composition is used to treat human CD38-related diseases.

In some embodiments, the CD38-associated disease includes multiple myeloma, non-Hodgkin's lymphoma, AL amyloidosis, T-cell lymphoma, diffuse large B-cell lymphoma, chronic lymphocytic leukemia, acute lymphoblastic cell leukemia, plasma cell leukemia, acute myelogenous leukemia, non-small cell lung cancer, pancreatic cancer, colorectal cancer and triple-negative breast cancer, etc.

In other aspects, the present application provides a method for preventing or treating human CD38-related diseases, comprising administering the antibody or antigen-binding portion thereof according to the first aspect, or the pharmaceutical composition according to the fifth aspect, to an individual in need.

The anti-human CD38 antibody or the antigen-binding part thereof of the present invention can specifically bind to human CD38, has high, medium and low affinity anti-human CD38 antibodies, and can kill tumor cells through an Fc-dependent immune effector mechanism. The anti-human CD38 antibody of the present invention or its antigen-binding portion can be used to prevent or treat CD38-related diseases, such as multiple myeloma, non-Hodgkin's lymphoma, AL-type amyloidosis, T-cell lymphoma and diffuse large B-cell lymphoma, etc.

Description of drawings

Figure 1 shows the experimental results of humanized CD38 monoclonal antibody binding to Daudi cells

Figure 2 shows the experimental results of humanized CD38 antibody-induced apoptosis of Daudi lymphoma cells

Figure 3 shows the experimental results of humanized CD38 antibody anti-Fc cross-linking induced apoptosis of Daudi lymphoma cells

Detailed ways

The present application provides novel anti-CD38 antibodies or antigen-binding portions thereof that specifically bind to human CD38. In a preferred embodiment, the antibody or antigen-binding portion thereof of the present application has high, medium and low affinity binding activities to CD38, and can also bind cynomolgus monkey CD38. The present application also provides the amino acid of the antibody or its antigen-binding fragment, the carrier comprising the amino acid, the host cell comprising the amino acid or the vector, the method for preparing and purifying the antibody, and the medical treatment of the antibody or its antigen-binding fragment. And biological applications, such as prevention or treatment of CD38-related diseases or conditions.

To facilitate understanding of this application, certain terms used herein are first defined.

The term "antibody" as used herein refers to an immunoglobulin molecule comprising four polypeptide chains, two heavy (H) chains and two light (L) chains interconnected by disulfide bonds, as well as multimers thereof (such as IgM ). Each heavy chain is comprised of a heavy chain variable region (abbreviated VH) and a heavy chain constant region (abbreviated CH). The heavy chain constant region comprises three domains, CH1, CH2 and CH3. Each light chain is comprised of a light chain variable region (abbreviated VL) and a light chain constant region (abbreviated CL). The light chain constant region comprises one domain (CL1). The VH and VL regions can be further subdivided into hypervariable regions called complementarity determining regions (CDRs), interspersed with conserved regions called framework regions (FRs).

As used herein, the term "antigen-binding portion" of an antibody refers to a portion or segment of an intact antibody molecule that is responsible for binding antigen. The antigen binding domain may comprise a heavy chain variable region (VH), a light chain variable region (VL), or both. Antigen-binding fragments of antibodies can be prepared from intact antibody molecules using any suitable standard technique, including proteolytic digestion or recombinant genetic engineering techniques, among others. Non-limiting examples of antigen binding portions include: Fab fragments; F(ab')2 fragments; Fd fragments; Fv fragments; single chain Fv (scFv) molecules; The smallest unit of recognition composed of residues (eg, isolated CDRs). The term "antigen-binding portion" also includes other engineered molecules, such as diabodies, triabodies, tetrabodies, and minibodies.

As used herein, the terms "heavy chain variable region (VH)" and "light chain variable region (VL)" refer to a single antibody variable heavy and light chain region, respectively, comprising FR1, 2, 3, and 4 and the CDRs 1, 2 and 3.

It is well known to those skilled in the art that complementarity determining regions (CDRs, usually CDR1, CDR2 and CDR3) are regions in the variable region that have the greatest influence on the affinity and specificity of antibodies. There are two common ways to define the CDR sequence of VH or VL, namely kabat definition and Chothia definition, see for example Kabat et al, "Sequences of Proteins of Immunological Interest", National Institutes of Health, Bethesda, Md. (1991); A1- Lazikani et al., J. Mol. Biol. 273:927-948 (1997); and Martin et al., Proc. Natl. Acad. Sci. USA 86:9268-9272 (1989). For the variable region sequence of a given antibody, the CDR region sequences in the VH and VL sequences can be determined according to the Kabat definition or the Chothia definition. In an embodiment of the present application, Kabat is used to define CDR sequences. Herein, CDR1, CDR2 and CDR3 of the heavy chain variable region are abbreviated as HCDR1, HCDR2 and HCDR3, respectively; CDR1, CDR2 and CDR3 of the light chain variable region are abbreviated as LCDR1, LCDR2 and LCDR3, respectively.

For the variable region sequence of a given antibody, the CDR region sequence in the variable region sequence can be analyzed in various ways, for example, it can be determined using the online software Abysis (http://www.abysis.org/).

The term "specific binding" as used herein refers to a non-random binding reaction between two molecules, for example the binding of an antibody to an antigenic epitope, for example an antibody with an affinity at least two times greater than its affinity for a non-specific antigen The ability to bind with affinity to a specific antigen. It is understood, however, that an antibody is capable of specifically binding to two or more antigens that are related in sequence. For example, antibodies of the invention can specifically bind to CD38 in humans and non-humans (eg, mice or non-human primates).

The term "monoclonal antibody" as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, ie, the individual antibodies comprising the population are identical except for the possible presence of naturally occurring mutations in a small number of individuals. The monoclonal antibodies described herein specifically include "chimeric" antibodies, in which a portion of the heavy and/or light chain is identical or homologous to the corresponding sequence in an antibody derived from a particular species or belonging to a particular antibody class or subclass, and the heavy chain and/or the remainder of the light chain is identical or homologous to the corresponding sequence in an antibody derived from another species or belonging to another antibody class or subclass, and also includes fragments of such antibodies, so long as they exhibit the desired (See, US Pat. No. 4,816,567; and Morrison et al, Proc. Natl. Acad. Sci. USA 81:6851-6855 (1984)).

As used herein, the term "homology" is defined as the percentage of residues in amino acid or nucleotide sequence variants that are identical after alignment of the sequences and introduction of gaps, if desired, to achieve a maximum percentage homology. Methods and computer programs for alignment are well known in the art. The "at least 80% homology" mentioned herein refers to any value from 80% to 100% homology, such as 85%, 90%, 95%, 99% and so on.

As used herein, the term "CD38-associated disease" includes multiple myeloma, non-Hodgkin's lymphoma, AL amyloidosis, T-cell lymphoma, diffuse large B-cell lymphoma, and the like.

In one aspect, the application provides an antibody or antigen-binding portion thereof that specifically binds CD38, comprising a heavy chain variable region and/or a light chain variable region. Tables 1-3 exemplarily list the CDR, heavy chain and light chain variable region amino acid sequences suitable for the antibodies disclosed in the present application.

In specific embodiments, HCDR3 is selected from the amino acid sequences shown in SEQ ID NOs: 3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, 39, 42, 45 and 48 . In another specific embodiment, HCDR3 is selected from the amino acid sequences shown in SEQ ID NOs: 3, 9, 18, 21, 27, 33, 36 and 45. In a preferred embodiment, HCDR3 is selected from the amino acid sequences shown in 9, 18, 21, 27, 33, 36 and 45.

In specific embodiments, HCDR2 is selected from the amino acid sequences shown in SEQ ID NOs: 2, 5, 8, 11, 14, 17, 20, 23, 26, 29, 32, 35, 38, 41, 44 and 47 . In another specific embodiment, HCDR2 is selected from the amino acid sequences shown in SEQ ID NOs: 2, 8, 17, 20, 26, 32, 35 and 44. In a preferred embodiment, HCDR2 is selected from the amino acid sequences shown in 8, 17, 20, 26, 32, 35 and 44.

In specific embodiments, HCDR1 is selected from the amino acid sequences shown in SEQ ID NOs: 1, 4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40, 43 and 46 . In another specific embodiment, HCDR1 is selected from the amino acid sequences shown in SEQ ID NOs: 1, 7, 16, 19, 25, 31, 34 and 43. In a preferred embodiment, HCDR1 is selected from the amino acid sequences shown in 7, 16, 19, 25, 31, 34 and 43.

In some embodiments, an antibody heavy chain variable region disclosed herein comprises an amino acid sequence selected from the group consisting of SEQ ID NOs:97, 99, 101, 103, 105, 107, 109, and 111. In specific embodiments, said heavy chain variable region consists of an amino acid sequence selected from the group consisting of SEQ ID NOs:97, 99, 101, 103, 105, 107, 109 and 111.

An antibody or antigen-binding portion thereof disclosed herein may further comprise a light chain variable region in addition to a heavy chain variable region.

In some embodiments, the CDR3 (LCDR3) of the light chain variable region is selected from SEQ ID NOs: 51, 54, 57, 60, 63, 66, 69, 72, 75, 78, 81, 84, 87, Amino acid sequences shown at 90, 93, 96 and 117. In another specific embodiment, LCDR3 is selected from the amino acid sequences shown in SEQ ID NOs: 51, 57, 66, 69, 75, 81, 84 and 117. In a preferred embodiment, LCDR3 is selected from the amino acid sequences shown in SEQ ID NOs: 57, 66, 69, 75, 81, 84 and 117.

In some embodiments, LCDR2 is selected from the amino acids shown in SEQ ID NOs: 50, 53, 56, 59, 62, 65, 68, 71, 74, 77, 80, 83, 86, 89, 92, 95 and 116 sequence, or selected from the amino acid sequences shown in SEQ ID NOs: 50, 65, 68, 74, 80, 83, 92 and 116. In a preferred embodiment, LCDR2 is selected from the amino acid sequences shown in SEQ ID NOs: 65, 68, 74, 80, 83, 92 and 116.

In some embodiments, LCDR1 is selected from the amino acid sequence of SEQ ID NOs: 49, 52, 55, 58, 61, 64, 67, 70, 73, 76, 79, 82, 85, 88, 91, 94, and 115, Or an amino acid sequence selected from SEQ ID NOs: 55, 64, 67, 73, 79, 82, 91 and 115. In a preferred embodiment, LCDR1 is selected from the amino acid sequences shown in SEQ ID NOs: 55, 64, 67, 73, 79, 82 and 91.

In some embodiments, an antibody light chain variable region disclosed herein comprises an amino acid sequence selected from the group consisting of SEQ ID NOs:98, 100, 102, 104, 106, 108, 110, and 112. In specific embodiments, said light chain variable region consists of an amino acid sequence selected from the group consisting of SEQ ID NOs:98, 100, 102, 104, 106, 108, 110 and 112.

In some embodiments, the heavy chain or the heavy chain variable region, the light chain or the light chain variable region of the antibody disclosed herein can be substituted, deleted or added at least one on the basis of the respective specific amino acid sequences listed above Amino acids, and the resulting variant still retains the activity of binding human CD38.

In certain embodiments, the number of amino acid substitutions, deletions or additions is 1-30, preferably 1-20, more preferably 1-10. In preferred embodiments, sequence variants differ from the original amino acid sequence by about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions, deletions, and/or additions. In more preferred embodiments, the sequence variant differs from the original amino acid sequence by a substitution, deletion or addition of about 1, 2, 3, 4 or 5 amino acids. In specific embodiments, the amino acid substitutions are conservative substitutions.

In a preferred embodiment, the antibody disclosed herein is antibody 57H, 105H, 116H, or 145H, wherein the heavy chain variable region sequence of antibody 57H is shown in SEQ ID NO: 99, and the light chain variable region sequence is shown in SEQ ID NO: 100, wherein LCDR2 is preferably shown in SEQ ID NO: 116, and the rest of the CDR sequence is the same as that of murine antibody 57; the heavy chain variable region sequence of antibody 105H is shown in SEQ ID NO: 107, and the light chain can be The variable region sequence is shown in SEQ ID NO: 108, wherein the CDR sequence is the same as that of murine antibody 105; the heavy chain variable region sequence of antibody 116H is shown in SEQ ID NO: 109, and the light chain variable region sequence is shown in SEQ ID NO: 110, wherein the CDR sequence is the same as that of murine antibody 116; the heavy chain variable region sequence of antibody 145H is shown in SEQ ID NO: 111, and the light chain variable region sequence is shown in SEQ ID NO: 112, wherein LCDR3 is preferably Shown in SEQ ID NO: 117, the rest of the CDR sequence is the same as that of murine antibody 145.

In some embodiments, the antibodies disclosed herein are monoclonal antibodies. In specific embodiments, the antibodies disclosed herein are humanized antibodies.

Antibodies disclosed herein, or antigen-binding portions thereof, are capable of specifically binding human CD38. In specific embodiments, the antibody, or antigen-binding portion thereof, specifically binds human CD38 or cynomolgus CD38. In preferred embodiments, the antibody or antigen binding portion thereof specifically binds human CD38.

In some embodiments, any suitable expression vector may be used in the present application. For example, the expression vector may be one of pTT5, pUC57, pDR1, pcDNA3.1(+), pDHFF and pCHO 1.0. Expression vectors may include fusion DNA sequences linked with appropriate transcriptional and translational regulatory sequences.

In some embodiments, the available host cells are cells containing the above-mentioned expression vectors, which may be eukaryotic cells, such as mammalian or insect host cell culture systems, which can be used for the expression of the antibody or its antigen-binding portion of the present application. For example, HEK293 cells, COS, CHO, NSO, sf9, and sf21 are all applicable to the present invention. The host cell may also be a prokaryotic cell containing the above expression vector, such as DH5α, BL21(DE3) or TG1, etc.

In some embodiments, the preparation method of the anti-human CD38 monoclonal antibody disclosed herein comprises: culturing host cells under suitable expression conditions, thereby expressing the anti-human CD38 monoclonal antibody; isolating and purifying the expressed anti-human CD38 monoclonal antibody Antibody. Using the above method, the recombinant protein can be purified into a substantially uniform substance, such as a single band on SDS-PAGE electrophoresis.

In some embodiments, the anti-human CD38 antibody disclosed herein can be separated and purified by using affinity chromatography. According to the characteristics of the affinity column used, conventional methods such as high-salt buffer, changing pH, etc. can be used. Methods Elution of anti-human CD38 antibody bound to the affinity column.

In some embodiments, the humanized anti-human CD38 monoclonal antibody disclosed herein is obtained by the following method: the human CD38 antigen prepared in the laboratory is used to immunize Balb/c mice, and the titer of the mice after multiple immunizations is compared with Then take mouse splenocytes to fuse with hybridoma cells and screen out hybridoma cell lines with different affinity. More specifically, through a large number of experiments, the inventors of the present application firstly expressed human CD38 antigen respectively, and on this basis, used different adjuvants to mix with human CD38 antigen to immunize mice, and then further hybridized the splenocytes of the above mice with The tumor cell line sp2/0 was fused, and the fused hybridoma was screened by human CD38 antigen to select a positive cell line, and the target cell line was obtained after verifying its binding to human CD38 and Daudi cells. After humanizing the target molecule, the light chain and heavy chain genes were simultaneously cloned into the eukaryotic expression vector pCHO1.0. The above expression vector was transfected into CHO cells by liposome method, and then the positive cell clones were screened with puromycin and methotrexate, and the screened high-expression clones were expanded and cultured in serum-free medium, and protein A affinity column was used to Isolation or purification of humanized anti-human CD38 antigen monoclonal antibody.

In other embodiments, conventional techniques in the art, such as PCR mutagenesis, can be used to further alter the murine parental antibody to generate chimeric or humanized or other variant forms of the antibody. The parent antibody of the present application can be mutated, for example, within the complementarity determining region (CDR) domain to generate a variant antibody that can be screened for the presence of desired properties, such as binding affinity (lower KD), IC50, specificity, preferential binding, etc. wait. Preferably, the property of interest in the variant antibody is an improvement over the property in the parental antibody. Amino acid substitution variant antibodies are preferred, and at least 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acid residues of the parent antibody molecule are removed and a different residue is inserted in its place. The sites of most interest for substitution mutagenesis are one or more CDR regions, but framework region (FR) alterations are also considered. Conservative amino acid substitutions are preferred, and non-conservative amino acid changes can also be introduced and the obtained variant antibodies can be used to screen for the desired properties.

The present application provides a pharmaceutical composition comprising an antibody disclosed herein, or an antigen-binding portion thereof, and a pharmaceutically acceptable carrier. The anti-human CD38 monoclonal antibody disclosed herein can be formulated into a pharmaceutical preparation together with a pharmaceutically acceptable carrier, so as to exert a more stable curative effect. In some embodiments, these formulations can ensure the conformational integrity of the amino acid core sequence of the anti-human CD38 monoclonal antibodies disclosed herein, while also protecting the multifunctional groups of the protein from degradation (including but not limited to aggregation, deamidation, or oxidation) . In some embodiments, liquid formulations are usually stable for at least one year when stored at 2°C-8°C. In some embodiments, for lyophilized formulations, it is stable for at least six months at 30°C.

In some embodiments, the anti-human CD38 antibody monoclonal antibody preparations can be suspension, aqueous injection, freeze-dried and other preparations commonly used in the pharmaceutical field, preferably aqueous injection or freeze-dried preparations, for the anti-human CD38 monoclonal antibody disclosed herein For the aqueous injection or freeze-dried preparation of the antibody, the pharmaceutically acceptable excipients include but are not limited to: surfactants, solution stabilizers, isotonic regulators, buffers or combinations thereof. In some embodiments, surfactants include, but are not limited to: nonionic surfactants such as polyoxyethylene sorbitan fatty acid esters (Tween 20 or 80), poloxamers (such as poloxamer 188), Triton, sodium dodecyl sulfate (SDS), sodium lauryl sulfate, tetradecyl, linoleyl or octadecyl sarcosine, Pluronics, MONAQUATTM, etc., the amount added should make the anti-human CD38 monoclonal antibody The tendency to graininess is minimal. In some embodiments, solution stabilizers include, but are not limited to, one or a combination of the following: sugars, such as reducing sugars and non-reducing sugars; amino acids, such as monosodium glutamate or histidine; alcohols Classes, such as trihydric alcohols, higher sugar alcohols, propylene glycol, polyethylene glycol, etc. The solution stabilizer is added in such an amount that the resulting formulation remains stable for a period of time considered stable by those skilled in the art. Isotonicity adjusting agents include, but are not limited to, sodium chloride, mannitol, or combinations thereof. Buffers include, but are not limited to: Tris, histidine buffer, phosphate buffer, or combinations thereof.

The present application also provides a method for preventing or treating CD38-related diseases, which comprises administering an anti-human CD38 antibody or a composition comprising an anti-human CD38 antibody to an individual. Specifically, the anti-human CD38 antibodies disclosed herein can effectively prevent and/or treat CD38-related diseases.

In some embodiments, the CD38-associated disease includes multiple myeloma, non-Hodgkin's lymphoma, AL amyloidosis, T-cell lymphoma, diffuse large B-cell lymphoma, chronic lymphocytic leukemia, acute lymphoblastic cell leukemia, plasma cell leukemia, acute myelogenous leukemia, non-small cell lung cancer, pancreatic cancer, colorectal cancer and triple-negative breast cancer, etc.

When the anti-human CD38 monoclonal antibody disclosed herein and the composition thereof are administered to animals including humans, the dosage varies according to the age and weight of the patient, the nature and severity of the disease, and the route of administration. Reference can be made to the Based on the results of the experiment and the overall situation, the total dosage should not exceed a certain range.

The dosage and frequency of administration of an antibody or composition thereof may vary depending on whether the disease is being prevented or treated. In prophylactic applications, the composition containing the antibody of the present application or its mixture is administered to patients who are not yet in a disease state to enhance the patient's resistance, and this amount is defined as "prophylactic effective dose". In this application, the specific dose depends on the patient's health status and systemic immunity. Generally, relatively lower doses are administered at relatively infrequent intervals for a longer period of time. In therapeutic applications, it is sometimes desirable to administer relatively higher doses at relatively shorter intervals until disease progression is slowed or terminated, and preferably until the patient shows partial or complete amelioration of disease symptoms. Thereafter, prophylactic regimens can be administered to the patient. Those of ordinary skill in the art can easily grasp the specific dosage and frequency according to actual needs.

In the present description and claims, the words "comprising", "comprising" and "containing" mean "including but not limited to", and are not intended to exclude other parts, additives, components, or steps.

It should be understood that features, characteristics, components or steps described in one particular aspect, embodiment or example of the present application can be applied to any other aspect, embodiment or example described herein unless incompatible therewith.

The above disclosure generally describes the present invention. The following specific examples are to further illustrate the present invention, and should not be construed as limiting the present invention. The examples do not include detailed descriptions of traditional routine methods, which are well known to those of ordinary skill in the art and are described in numerous publications, for example Handbook of Molecular Cloning, Laboratory Handbook of Antibody Technology of Cold Spring Harbor . Reagent sources not indicated are conventional reagents.

specific embodiment

Example 1 Preparation of the Fc tag and Flag tag antigen of the extracellular region of human CD38, the Flag tag protein of the extracellular region of cynomolgus monkey CD38 , and the reference antibody Daratumumab

The human CD38 antigen sequence was purchased from Sino Biological (Product No.: HG10818-M). The 43-300th amino acid fragment of the N-terminal of human CD38 was spliced with the hFc fragment and the Flag tag (DYKDDDDK) by the PCR method, and the hFc fragment and the Flag tag were located at the C-terminal, and constructed on the pTT5 expression vector (preserved in the laboratory) to obtain For pTT5(hCD38-ECD-hFc) and pTT5(hCD38-ECD-Flag), clones with completely correct sequencing results were selected for plasmid extraction and transfection.

The cynomolgus monkey CD38 sequence was purchased from Sino Biological (Cat. No.: CG90050-G). The 44th-301th amino acid fragment of its N-terminal was spliced with the Flag tag (DYKDDDDK) by PCR method, and the Flag tag was located at the C-terminal, and it was constructed on the pTT5 expression vector (preserved in the laboratory) to obtain pTT5 (cyno-CD38-ECD -Flag), select clones with completely correct sequencing results for plasmid extraction and transfection.

The amino acid sequence of Daratumumab was obtained from IMGT, and the nucleotide sequences of the variable regions of the heavy and light chains were synthesized from the whole gene after codon optimization. The variable region of the heavy chain of Daratumumab was connected to the constant region of IgG1 by PCR; the variable region of the light chain of Daratumumab was connected to the constant region of the Kappa chain. These fragments were then cloned into the pTT5 expression vector, and plasmid extraction was carried out after sequencing verification to prepare for transfection.

The plasmid was transfected into HEK293E cell line (preserved in laboratory) by PEI method. After culturing for 5 days in Freestyle293 medium (purchased from Gibco) containing 3 mM valproic acid, protein A affinity chromatography (purchased from Pharmacia) or Flag affinity chromatography (purchased from Genscript) Purify the protein of interest from the serum. The protein was quantified by bicinchoninic acid (BCA) method, and the purified protein was used for the following further analysis and research. The purified protein was used for the following mouse immunization and further analysis and research.

Example 2 Immunization of hCD38-ECD-hFc

Dilute 100 μg/mouse hCD38-ECD-hFc antigen with physiological saline to 75 μl, mix with an equal volume of Freund’s complete adjuvant, and complete phacoemulsification for 4-5 week-old Balb/c mice (purchased Subcutaneous multi-point injection was performed from Shanghai Lingchang Biotechnology Co., Ltd., animal production license number: SCXK (Shanghai) 2013-0018). Three weeks later, 50 μg/mouse protein was also diluted to 75 μl and mixed with an equal volume of Freund’s incomplete adjuvant. After phacoemulsification was complete, the mice were injected subcutaneously at multiple points, and the immunization was repeated two weeks later. One week after the third immunization, the tails of all mice were cut to collect blood to separate serum, and the serum titer was detected by ELISA coated with hCD38-ECD-hFc antigen. For mice with serum antibody titer > 10000, pulse immunization was performed one week after blood collection: 10 μg antigen/100 μl normal saline/mouse were injected into the tail vein.

The detection of the titer was carried out by ELISA method: the ELISA plate was coated with hCD38-ECD-hFc antigen, the coating concentration was 1 μg/ml, 100 μl per well, and coated overnight at 4°C. Plates were washed twice with PBST (PBS containing 0.5% Tween-20) and then patted dry. Add 200 μl of coating solution containing 1% BSA to each well to seal, block at room temperature for 4 hours, pat dry, and store in a -20°C refrigerator until use. During the detection, 100 μl of mouse serum of different concentrations was added to each well of the ELISA plate, and two duplicate wells were set up, and incubated at room temperature for 1.5 hours. After washing 3 times with PBST, pat dry. Add 100 μl of HRP-labeled rabbit anti-mouse Ig antibody (purchased from Sigma) diluted 1:10000 times with PBST, and incubate at room temperature for 1 hour. After washing 3 times with PBST, pat dry. Add 100 μl of chromogenic solution to each well (mix ELISA chromogenic solution A and chromogenic B solution at a volume ratio of 1:1 before use) for color development, and then add 100 μl of 2M H2SO4 stop solution to each well to stop the reaction. Immediately measure the OD value of each well with a microplate reader (Molecular Device) at a wavelength of 450 nm.

Embodiment 3 hybridoma fusion and screening

Hybridoma sp2/0 cells (from the Cell Bank of the Type Culture Collection Committee of the Chinese Academy of Sciences, with the preservation number TCM-18) were cultured in a 37°C, 5% CO ₂ incubator, and the medium was changed one day before fusion. Three days after the shock immunization of the mice, the splenocytes of the mice were taken for fusion. The fusion and screening method is as follows: the spleen of the mouse is taken, ground and washed, and the spleen cells are counted. Splenocytes and sp2/0 cells were mixed at a ratio of 10:1 and centrifuged at 1500rpm for 7 minutes. Wash away the supernatant. Add 1ml of PEG (1450) within 1 minute, shake gently for 90 seconds, add 5ml of serum-free DMEM culture solution (purchased from Gibco) within 2.5 minutes, and then add 5ml of serum-free culture solution to stop the reaction, and let it stand for 5 minutes. Centrifuge at 1280 rpm for 8 minutes. According to the number of 2 million sp2/0 cells in a 96-well plate, the cells were evenly seeded into a 96-well plate, 200 μl per well. First use HAT medium containing hypoxanthine (H), methotrexate (A) and thymidine (T) to screen, change half of the medium every 3 to 4 days, and change the medium on the 10th day. Use HT medium. After 10 days, when the hybridoma cells covered more than 10% of the bottom of the 96-well plate, the supernatant was taken for ELISA detection with an enzyme label plate coated with hCD38-ECD-hFc antigen. The ELISA detection method is the same as that described in Example 2. The positive hybridoma clones were selected and cultured in a 24-well plate, subcloned by the limiting dilution method, and the hybridoma strains stably expressing the target antibody were obtained to preserve the species and build a library.

Determination of the sequence of the anti-human CD38 monoclonal antibody of embodiment 4 mouse origin

After 2-3 rounds of subcloning for the positive wells with higher affinity, 150 hybridoma strains were finally obtained. Antibody sequences were retrieved from these hybridoma cell lines. The total RNA of each hybridoma cell line was extracted using Trizol, and the mRNA was reverse-transcribed into cDNA using a reverse transcription kit (purchased from ABI). The light chain variable region and heavy chain variable region genes of the mouse-derived anti-human CD38 monoclonal antibody were amplified by PCR with primers reported in the literature, and then the PCR product was cloned into pGEM-T vector, sequenced and analyzed for the variable region genes sequence. The obtained sequences were compared and analyzed in GenBank, and all the sequences conformed to the characteristics of mouse IgG variable region genes. Table 1 and Table 2 list the amino acid sequences of the CDR regions of preferred antibodies.

Table 1: Amino acid sequences of heavy chain CDRs of exemplary murine anti-human CD38 antibodies

Example 5 Humanization of anti-human CD38 monoclonal antibody

According to the sequence analysis results, antibodies 1, 57, 71, 73, 77, 78, 105, 116 and 145 were selected to construct chimeric antibodies and humanized antibodies. The chimeric antibody is constructed by truncating the heavy chain variable region and light chain variable region of the murine antibody, and connecting them with the light and heavy chain constant regions of human IgG1 respectively by overlapping PCR.

Table 2: Light chain CDR amino acid sequences of exemplary murine anti-human CD38 antibodies

According to the Kabat rule, the amino acid sequences of the light chain variable region and the heavy chain variable region of the mouse anti-human CD38 monoclonal antibody were analyzed and 3 CDRs and 4 FRs were determined. Taking antibody No. 105 as an example, IGHV1-46*01 was selected as the heavy chain CDR grafting template by comparing the homology between NCBI IgBlast and human IgG germline sequence (Germline), and the mouse-derived anti-human CD38 monoclonal antibody 105 The heavy chain CDR region of No. 1 was grafted into the framework region of IGHV1-46*01 to construct a heavy chain CDR grafted antibody. Similarly, after comparing the homology with the human IgG germline sequence, IGKV1-16*01 was selected as the light chain CDR transplantation template, and the light chain CDR region of the mouse-derived anti-human CD38 monoclonal antibody No. 105 was transplanted into IGKV1-16 * The skeleton region of 01 is constructed as a light chain CDR-grafted antibody. At the same time, on this basis, some amino acid positions in the framework region were back-mutated. When performing back mutation, the amino acid sequence was coded by Kabat, and the position of the site was indicated by the Kabat code. Preferably, for the light chain variable region sequence, the F at position 36 of the Kabat code is restored to L of mouse origin, the A at position 43 is mutated to T, the P at position 44 is restored to I of mouse origin, and the The S at position 46 was converted to the mouse-derived R, the G at position 66 was converted to the mouse-derived R, and the T at position 69 was converted to the mouse-derived S. For the heavy chain variable region sequence, the M at position 48 of the Kabat code is restored to I of murine origin, the V at position 67 is restored to A of murine origin, the M at position 69 is restored to L of murine origin, and the 71st position The R at position 73 is restored to mouse-derived A, the T at position 73 is restored to mouse-derived K, and the V at position 78 is restored to mouse-derived A. The above variable region gene sequence was codon-optimized and synthesized by Sangon Biotech according to the codon usage preference of Cricetulus griseus. The synthetic humanized variable region sequence was connected with the human IgG1 constant region, and this antibody was defined as the humanized antibody 105-Humanization of antibody 105, 105H).

Using the same principle as above, the remaining antibodies were similarly humanized. The amino acid sequence number of the variable region of the humanized antibody is shown in Table 3, the amino acid sequence of the heavy chain constant region is shown in SEQ ID NO: 113, and the amino acid sequence of the light chain constant region is shown in SEQ ID NO: 114. The pTT5 vector was used to construct the transient expression vectors of the humanized heavy chain and light chain respectively, and the combination of the above light and heavy chains was transiently transfected and expressed antibodies using the HEK293E system. HEK293E cells were cultured in Free Style293Expression Medium (purchased from Gibco) medium, and the plasmid was transferred into the cells by PEI transfection method 5 days later, the cell supernatant was collected, purified by Protein A, and each humanized monoclonal antibody was obtained.

Finally, the amino acid sequence of the heavy chain variable region of antibody No. 1 after humanization is shown in SEQ ID NO:97; the amino acid sequence of the humanized light chain variable region is shown in SEQ ID NO:98. After linking with the constant region of human IgG1, the antibody sequence of 1H is finally obtained.

The amino acid sequence of the heavy chain variable region of antibody No. 57 after humanization is shown in SEQ ID NO: 99; the amino acid sequence of the humanized light chain variable region is shown in SEQ ID NO: 100. After linking with the constant region of human IgG1, the antibody sequence of 57H is finally obtained.

The amino acid sequence of the heavy chain variable region of antibody No. 71 after humanization is shown in SEQ ID NO: 101; the amino acid sequence of the humanized light chain variable region is shown in SEQ ID NO: 102. After linking with the constant region of human IgG1, the antibody sequence of 71H was finally obtained.

The amino acid sequence of the humanized heavy chain variable region of antibody No. 73 is shown in SEQ ID NO: 103; the amino acid sequence of the humanized light chain variable region is shown in SEQ ID NO: 104. After linking with the constant region of human IgG1, the antibody sequence of 73H was finally obtained.

The amino acid sequence of the heavy chain variable region after humanization of antibody No. 78 is shown in SEQ ID NO: 105; the amino acid sequence of the humanized light chain variable region is shown in SEQ ID NO: 106. After linking with the constant region of human IgG1, the antibody sequence of 78H is finally obtained.

The amino acid sequence of the heavy chain variable region of antibody No. 105 after humanization is shown in SEQ ID NO: 107; the amino acid sequence of the humanized light chain variable region is shown in SEQ ID NO: 108. After linking with the constant region of human IgG1, the antibody sequence of 105H is finally obtained.

The amino acid sequence of the heavy chain variable region of antibody No. 116 after humanization is shown in SEQ ID NO: 109; the amino acid sequence of the humanized light chain variable region is shown in SEQ ID NO: 110. After linking with the human IgG1 constant region, the antibody sequence of 116H was finally obtained.

The amino acid sequence of the heavy chain variable region of antibody No. 145 after humanization is shown in SEQ ID NO: 111; the amino acid sequence of the humanized light chain variable region is shown in SEQ ID NO: 112. After linking with the constant region of human IgG1, the antibody sequence of 145H is finally obtained.

Table 3: Amino acid sequences of heavy and light chain variable regions of exemplary humanized anti-human CD38 antibodies

Example 6 Binding of humanized CD38 monoclonal antibody to Daudi cells

The well-growing Daudi cells (preserved in the laboratory) were sampled and counted, centrifuged and resuspended in PBS supplemented with 1% FBS, and the density was adjusted to 2X 10 ⁷ . The highest concentration of the antibody sample is 10 μg/ml, and the sample is added after three-fold serial dilution (8 gradients in total). In a 96-well culture plate, 20 ul/well (4X 10 ⁵ /well) Daudi cells were added, and 30 μl of gradiently diluted antibody drug was added, and incubated at 4°C for 30 minutes. Add 180 μl of 1% FBS and PBS to wash, and centrifuge at 1200 rpm for 6 min. After the supernatant was sucked off with a vacuum pump, 50 μl of goat anti-human IgG-FITC (diluted with 1% FBS and PBS 1:1000) was added, and incubated at 4° C. for 30 minutes. After adding 180 μl of 1% FBSPBS, centrifuge at 1200 rpm for 6 minutes and resuspend with 160 μl of PBS. Finally, the fluorescence intensity on the cell surface was detected by flow cytometry. Data analysis was performed using GraphPad Prism6 software. The experimental results are shown in Figure 1 and Table 4.

Table 4: Binding of humanized CD38 antibodies to Daudi cells

Example 7 Humanized CD38 Antibody Induces Daudi Lymphoma Cell Apoptosis

Add 1X 10 ⁵ Daudi cells/well, and 1.85 and 1.03 μg/well of antibody drugs in the experimental system of 200 μl/well in a 96-well culture plate. After 24 hours of culture, directly add 1:100 PI (Propidium Iodide) solution (2μl), continue to incubate for 30 minutes, centrifuge to remove the supernatant, wash with PBS, centrifuge to remove the supernatant, then add 160μl PBS to resuspend, and complete the detection with a flow cytometer. Set unstained wells as a blank control, and the reference antibody is Daratumumab (abbreviated as Dara in Figure 2). The results in Figure 2 show that 73H has stronger apoptosis-inducing activity than the reference antibody, and the other antibodies are consistent with the reference antibody, and no obvious apoptosis-inducing activity is seen.

Example 8 Humanized CD38 Antibody Anti-Fc Crosslinking Induces Daudi Lymphoma Cell Apoptosis

In the experimental system of 200 μl/well of 96-well culture plate, add 1X 105 Daudi cells/well, and add 1.85 and 1.03 μg/well of antibody drugs, and incubate in the cell culture incubator for 30 minutes. Then 1 μg/well anti-human Fc antibody was added, and culture was continued for 24 hours. Incubate for 24 hours. After the culture is over, directly add 1:100 PI solution (2μl) to the culture well, continue to culture for 30 minutes, centrifuge to remove the supernatant, wash with PBS, centrifuge to remove the supernatant, then add 160μl PBS to resuspend, and use a flow cytometer Complete the test. Set unstained wells as a blank control, and the reference antibody is Daratumumab (abbreviated as Dara in Figure 3). The results in Figure 3 show that the five preferred antibodies have consistently stronger anti-Fc cross-linking-induced apoptosis activity than the reference antibody.

Example 9 Affinity Determination of Humanized CD38 Monoclonal Antibody

The affinity of the expressed and purified humanized antibody was detected by Biacore T200 (GE healthcare). The specific experimental method is: using Protein-A CM5 sensor chip (GE healthcare), using FC1 (Flow cell 1) as the reference channel, and FC2 (Flow cell 2) as the sample channel. Human antibodies or control antibodies were captured in the FC2 channel, followed by injection of different concentrations of hCD38-Flag or cyno-CD38-Flag. The cycle conditions were as follows: inject analyte in all channels of FCs at 50 μl/min for 4 min, dissociate time for 20 min, inject 6M guanidine hydrochloride (Sinopharm Chemical Reagent Co., Ltd.) at a rate of 10 μl/min for 30 s for surface regeneration, and then use Biacore T200 Evaluation Software Ver 1.0 calculates the difference between the signal of the capture antibody and the signal without the capture antibody and the affinity of the interaction. As shown in Table 5 and Table 6. Experimental results show that the preferred antibodies can all bind to CD38 of humans and cynomolgus monkeys, with high, medium and low affinities.

Table 5: Binding affinity of humanized CD38 antibodies to human CD38

Antibody number Ka(1/Ms) Kd(1/s) KD(nM) 57H 2.641E+05 1.009E-04 0.382 105H 2.976E+04 9.748E-04 32.755 116H 1.472E+06 9.530E-05 0.065 145H 6.125E+05 1.975E-03 3.224

Table 6: Binding affinity of humanized CD38 antibodies to monkey CD38

Antibody number Ka(1/Ms) Kd(1/s) KD(nM) 57H 5.492E+04 9.201E-05 1.675 105H 6.113E+04 2.080E-04 3.402 116H 2.513E+05 1.019E-04 0.405 145H 9.782E+04 3.930E-03 40.176

Conclusion: The inventors of the present invention have conducted a large number of experiments and obtained a group of monoclonal antibodies that can specifically bind to CD38 on the cell surface, and these antibodies can also bind to CD38 in cynomolgus monkeys. A series of anti-human CD38 antibodies with high, medium and low affinity were obtained, which are able to kill tumor cells through Fc-dependent immune effector mechanisms.

It is to be understood that although the application has been illustrated in some form, the application is not limited to what is shown and described in this specification. It will be obvious to those skilled in the art that various changes may be made to the described embodiments and/or a certain feature or parameter without departing from the scope of the present application. These changes are all within the protection scope of this application.

SEQUENCE LISTING

<110> Shanghai Maiji Biotechnology Co., Ltd.

<120> Anti-human CD38 antibody, preparation method and use thereof

<130> anti-CD38

<160> 114

<170> PatentIn version 3.5

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<213> Mus musculus

<400> 74

Tyr Thr Ser Arg Leu His Ser

1 5

<210> 75

<211> 9

<212> PRT

<213> Mus musculus

<400> 75

Gln Gln Gly Asn Thr Leu Pro Phe Thr

1 5

<210> 76

<211> 16

<212> PRT

<213> Mus musculus

<400> 76

Arg Ser Ser Gln Ser Leu Val His Ser Asn Gly Asn Thr Tyr Leu His

1 5 10 15

<210> 77

<211> 7

<212> PRT

<213> Mus musculus

<400> 77

Lys Val Ser Asn Arg Phe Ser

1 5

<210> 78

<211> 9

<212> PRT

<213> Mus musculus

<400> 78

Ser Gln Ser Thr His Val Pro Pro Thr

1 5

<210> 79

<211> 11

<212> PRT

<213> Mus musculus

<400> 79

Arg Ala Ser Gln Glu Ile Ser Gly Tyr Leu Ser

1 5 10

<210> 80

<211> 7

<212> PRT

<213> Mus musculus

<400> 80

Ala Ala Ser Thr Leu Asp Ser

1 5

<210> 81

<211> 9

<212> PRT

<213> Mus musculus

<400> 81

Leu Gln Tyr Asp Ser Tyr Pro Leu Thr

1 5

<210> 82

<211> 11

<212> PRT

<213> Mus musculus

<400> 82

Lys Ala Ser Gln Asp Val Val Thr Thr Val Ala

1 5 10

<210> 83

<211> 7

<212> PRT

<213> Mus musculus

<400> 83

Ser Ala Ser Tyr Arg Tyr Ser

1 5

<210> 84

<211> 9

<212> PRT

<213> Mus musculus

<400> 84

Gln Gln Tyr Tyr Thr Tyr Pro Phe Thr

1 5

<210> 85

<211> 16

<212> PRT

<213> Mus musculus

<400> 85

Arg Ser Ser Gln Ser Ile Val His Ile Asn Gly Lys Thr Tyr Leu Glu

1 5 10 15

<210> 86

<211> 7

<212> PRT

<213> Mus musculus

<400> 86

Lys Ile Ser Asn Arg Phe Ser

1 5

<210> 87

<211> 9

<212> PRT

<213> Mus musculus

<400> 87

Phe Gln Gly Ser His Val Pro Arg Thr

1 5

<210> 88

<211> 16

<212> PRT

<213> Mus musculus

<400> 88

Arg Ser Ser Gln Ser Ile Val His Ile Asn Gly Lys Thr Tyr Leu Glu

1 5 10 15

<210> 89

<211> 6

<212> PRT

<213> Mus musculus

<400> 89

Lys Ile Ser Asn Arg Phe

1 5

<210> 90

<211> 9

<212> PRT

<213> Mus musculus

<400> 90

Phe Gln Gly Ser His Val Pro Arg Thr

1 5

<210> 91

<211> 11

<212> PRT

<213> Mus musculus

<400> 91

Lys Ala Ser Gln Asn Val Gly Thr Ser Val Ala

1 5 10

<210> 92

<211> 7

<212> PRT

<213> Mus musculus

<400> 92

Ser Thr Ser Tyr Arg Ser Ser

1 5

<210> 93

<211> 9

<212> PRT

<213> Mus musculus

<400> 93

Gln Gln Tyr Asn Ser Tyr Pro Leu Thr

1 5

<210> 94

<211> 11

<212> PRT

<213> Mus musculus

<400> 94

Arg Ala Ser Gln Asp Ile Ser Asn Tyr Leu Asn

1 5 10

<210> 95

<211> 7

<212> PRT

<213> Mus musculus

<400> 95

Tyr Thr Ser Arg Leu His Ser

1 5

<210> 96

<211> 9

<212> PRT

<213> Mus musculus

<400> 96

Gln Gln Gly Lys Thr Leu Pro Trp Thr

1 5

<210> 97

<211> 122

<212> PRT

<213> Synthetic

<400> 97

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Ser

20 25 30

Trp Met His Trp Ala Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile His Pro Asp Ser Gly Lys Thr Asn Tyr Asn Glu Lys Phe

50 55 60

Lys Gly Arg Ala Thr Leu Thr Val Asp Thr Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Tyr Leu Ser Ser Pro Tyr Tyr Tyr Val Met Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 98

<211> 112

<212> PRT

<213> Synthetic

<400> 98

Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser

20 25 30

Gly Gly Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Phe Cys Ser Gln Ser

85 90 95

Thr His Val Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 99

<211> 117

<212> PRT

<213> Synthetic

<400> 99

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser Phe

20 25 30

Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Tyr Ile Gly Gly Asp Ser Asn Ile Ile Tyr Tyr Ala Asp Thr Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Ser His Asp Tyr Thr Met Asp Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser

115

<210> 100

<211> 113

<212> PRT

<213> Synthetic

<400> 100

Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Val Thr Ile Thr Cys Lys Ser Ser Gln Ser Leu Leu Tyr Ser

20 25 30

Thr Asn Gln Lys Asn Tyr Leu Ala Trp Phe Gln Gln Lys Pro Gly Gln

35 40 45

Ser Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Arg

85 90 95

Tyr Tyr Asn Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys

<210> 101

<211> 123

<212> PRT

<213> Synthetic

<400> 101

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Lys Phe Thr Lys Tyr

20 25 30

Trp Ile Glu Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Ser Pro Gly Ile Asp Lys Thr Asn Tyr Asn Glu Lys Phe

50 55 60

Lys Asp Arg Ala Thr Phe Thr Ala Asp Thr Ser Thr Asn Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Glu Glu Tyr Gly Asn Ser Val Arg Tyr Ala Met Asp Tyr

100 105 110

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 102

<211> 107

<212> PRT

<213> Synthetic

<400> 102

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser Ser Val Ser Tyr Met

20 25 30

Tyr Trp Leu Gln Gln Lys Pro Gly Gln Ser Pro Arg Leu Leu Ile Tyr

35 40 45

Arg Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser

50 55 60

Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu

65 70 75 80

Asp Phe Ala Val Tyr Tyr Cys Gln Glu Arg Ser Ser Tyr Pro Pro Phe

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 103

<211> 119

<212> PRT

<213> Synthetic

<400> 103

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Gly Tyr

20 25 30

Asn Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Asn Ile Asp Pro Tyr Phe Gly Gly Thr Ile Tyr Asn Gln Lys Phe

50 55 60

Lys Gly Arg Val Thr Leu Thr Arg Asp Thr Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gln Tyr Gly Asn Trp Val Gly Phe Ala Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 104

<211> 107

<212> PRT

<213> Synthetic

<400> 104

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Val Thr Thr Ala

20 25 30

Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Ser Ala Ser Ser Arg Tyr Thr Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln His Phe Phe Phe Pro Tyr

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 105

<211> 121

<212> PRT

<213> Synthetic

<400> 105

Asp Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Tyr Ser Ile Ser Ser Ser Gly

20 25 30

Tyr Tyr Trp Asn Trp Phe Arg Gln Pro Pro Gly Lys Lys Leu Glu Trp

35 40 45

Ile Gly Tyr Arg Ser Tyr Asp Gly Ser Tyr Asn Tyr Asn Pro Ser Leu

50 55 60

Glu Asn Arg Ile Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Phe Ser

65 70 75 80

Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Glu Glu Arg Tyr Ile Leu Arg Tyr Ala Met Asp Tyr Trp Gly

100 105 110

Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 106

<211> 107

<212> PRT

<213> Synthetic

<400> 106

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Asn Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Thr Val Lys Leu Leu Ile

35 40 45

Tyr Tyr Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Gly Asn Thr Leu Pro Phe

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 107

<211> 120

<212> PRT

<213> Synthetic

<400> 107

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly His Ile Asn Pro Ser Ser Gly Tyr Ser Glu Tyr Ser Gln Lys Phe

50 55 60

Lys Asp Arg Ala Thr Leu Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Glu Ile Tyr Tyr Tyr Val Ser Arg Tyr Val Gly Asp Ser Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 108

<211> 107

<212> PRT

<213> Synthetic

<400> 108

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Glu Ile Ser Gly Tyr

20 25 30

Leu Ser Trp Leu Gln Gln Lys Pro Gly Lys Thr Ile Lys Arg Leu Ile

35 40 45

Tyr Ala Ala Ser Thr Leu Asp Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Arg Ser Gly Ser Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr Asp Ser Tyr Pro Leu

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 109

<211> 121

<212> PRT

<213> Synthetic

<400> 109

Gln Val Thr Leu Lys Glu Ser Gly Pro Ala Leu Val Lys Pro Thr Gln

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Leu Ser Gly Phe Ser Leu Ser Thr Ser

20 25 30

Arg Met Gly Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Leu Ala His Ile Trp Trp Asp Asp Asp Lys Arg Tyr Asn Pro Ala

50 55 60

Leu Lys Ser Arg Leu Thr Ile Ser Lys Asp Thr Ser Ser Asn Gln Val

65 70 75 80

Phe Leu Lys Ile Ala Ser Val Asp Lys Glu Asp Thr Ala Thr Tyr Tyr

85 90 95

Cys Gly Arg Ile Glu Val Gly Arg Gly Ala Trp Phe Ala Tyr Trp Gly

100 105 110

Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 110

<211> 107

<212> PRT

<213> Synthetic

<400> 110

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asp Val Val Thr Thr

20 25 30

Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Thr Leu Ile

35 40 45

Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Phe

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 111

<211> 115

<212> PRT

<213> Synthetic

<400> 111

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Leu Ile Asp Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Leu Ile Asn Pro Gly Ser Gly Ser Thr Lys Tyr Asn Glu Lys Phe

50 55 60

Lys Gly Arg Val Thr Leu Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Thr Gly Thr Leu Pro Tyr Trp Gly Gln Gly Thr Leu Val Thr

100 105 110

Val Ser Ser

115

<210> 112

<211> 107

<212> PRT

<213> Synthetic

<400> 112

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Ser

20 25 30

Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Ser Thr Ser Tyr Arg Ser Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Gly Ser Tyr Pro Leu

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 113

<211> 330

<212> PRT

<213> Homo sapiens

<400> 113

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu

225 230 235 240

Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

325 330

<210> 114

<211> 107

<212> PRT

<213> Homo sapiens

<400> 114

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

1 5 10 15

Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe

20 25 30

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

35 40 45

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

50 55 60

Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu

65 70 75 80

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

85 90 95

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

100 105

Claims (13)

1. An antibody, or antigen-binding portion thereof, that specifically binds human CD38,

the amino acid sequence of the HCDR1 of the antibody or antigen-binding portion thereof is set forth in SEQ ID NO:34, the amino acid sequence of HCDR2 is shown as SEQ ID NO:35, the amino acid sequence of HCDR3 is shown as SEQ ID NO:36, the amino acid sequence of LCDR1 is shown as SEQ ID NO:82, the amino acid sequence of LCDR2 is shown as SEQ ID NO:83, and the amino acid sequence of LCDR3 is shown in SEQ ID NO: as shown at 84.

2. The antibody, or antigen-binding portion thereof, of claim 1, wherein:

the amino acid sequence of the heavy chain variable region of the antibody is shown as SEQ ID NO:109, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 110.

3. The antibody or antigen-binding portion thereof of any one of claims 1-2, wherein the antigen-binding portion is selected from the group consisting of a Fab fragment, a Fab 'fragment, a F (ab') 2 fragment, a Fv fragment, and a scFv fragment.

4. The antibody or antigen-binding portion thereof of any one of claims 1-2, wherein the antibody is a murine monoclonal antibody or a humanized antibody.

5. A pharmaceutical composition comprising the antibody or antigen-binding portion thereof of any one of claims 1-4 and a pharmaceutically acceptable carrier.

6. An expression vector comprising a nucleotide molecule encoding the antibody or antigen-binding portion thereof of any one of claims 1-4.

7. The expression vector of claim 6, wherein the expression vector is pTT5, pUC57, pDR1, pcDNA3.1 (+), pDFHF or pCHO 1.0.

8. A host cell comprising the antibody or antigen-binding portion thereof of any one of claims 1-4, or comprising the expression vector of claim 6 or 7.

9. The host cell of claim 8, wherein the host cell is HEK293, COS, CHO, NS0, sf9, sf21, DH5 α, BL21 (DE 3), or TG1.

10. A method of producing the antibody, or antigen-binding portion thereof, of any one of claims 1-4, comprising:

a) Culturing the host cell of claim 8 or 9 under expression conditions such that the host cell is capable of producing the antibody, or antigen-binding portion thereof, thereby expressing the antibody, or antigen-binding portion thereof; and

b) Isolating and purifying the antibody or antigen-binding portion thereof expressed in step a).

11. Use of the antibody or antigen-binding portion thereof of any one of claims 1-4 or the pharmaceutical composition of claim 5 in the manufacture of a medicament for preventing or treating a CD 38-associated disease.

12. The use according to claim 11, wherein said CD38 related disease is selected from the group consisting of multiple myeloma, non-hodgkin's lymphoma, AL-type amyloidosis, T-cell lymphoma, diffuse large B-cell lymphoma, chronic lymphocytic leukemia, acute lymphocytic leukemia, plasma cell leukemia, acute myelogenous leukemia, non-small cell lung cancer, pancreatic cancer, colorectal cancer and triple negative breast cancer.

13. The use according to claim 11, wherein said CD38 associated disease is selected from the group consisting of multiple myeloma, non-hodgkin's lymphoma, AL-type amyloidosis, T-cell lymphoma, and diffuse large B-cell lymphoma.

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