CD38-targeted nano antibody, pharmaceutical composition and application thereof

Abstract

The invention provides a CD38-targeted nano antibody, a pharmaceutical composition and application of the CD38-targeted nano antibody, and relates to the technical field of biological medicines. The targeted CD38 nano antibody is any one of a first antibody and a second antibody and comprises three complementary determining regions; the sequences of the complementarity determining regions CDR1 to CDR2 of the first antibody are as shown in SEQ ID NO.1, SEQ ID NO.3 and SEQ ID NO.5; the sequences of the complementarity determining regions CDR1 to CDR2 of the second antibody are as shown in SEQ. ID NO.2, SEQ. ID NO.4 and SEQ. ID NO.6; the nano antibody has high affinity with purified and cell surface CD38 protein, and an effective novel method is provided for CD38 targeted anti-tumor intervention; and detection reagents such as molecular probes can be prepared on the basis of the nano antibody, and a new way is provided for tumor specific imaging and diagnosis through specific binding with the tumor surface CD38.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and more specifically to a CD38-targeting nano antibody, a pharmaceutical composition and applications thereof. Background Art

[0002] Tumor targeted therapy is a method of treating tumors by specifically targeting molecules or signaling pathways that play an important role in tumor occurrence and progression. Giving corresponding targeted therapy based on the specific molecular phenotype of the tumor has gradually become the preferred option for current tumor treatment. Tumor targeted drugs can be roughly divided into two categories: monoclonal antibodies and small molecule compounds. With the deepening of research, new targets continue to emerge, and anti-tumor drug research and development has made a number of breakthroughs. Tumor targeted drugs are increasing at a rate of 5 to 6 per year. However, the effectiveness of targeted therapy is limited by the heterogeneity of tumor cells, and different patients may have significant differences in their responses to the same drug; in addition, tumor cells may develop drug resistance through a variety of mechanisms, resulting in a decrease in the therapeutic effect. Therefore, for the time being, the development of targeted drugs is still insufficient, the widespread application of existing treatment methods is limited, and the vast majority of patients do not benefit from them. Therefore, there is an urgent need to discover new molecular targets and develop more effective molecular targeted therapy strategies to improve the accuracy and effectiveness of treatment and reduce the treatment burden on patients.

[0003] CD38 (Cluster of Differentiation 38) is a type II transmembrane glycoprotein with a molecular weight of approximately 46 kD. In 1980, CD38 was first discovered as a specific marker for T cell activation by scientists such as EL Reinherz and SF Schlossman. The CD38 protein sequence includes a short cytoplasmic tail at the N-terminus, a single transmembrane domain, and a longer extracellular region at the C-terminus. Current studies have shown that CD38 protein is a bifunctional extracellular enzyme with cyclase and hydrolase activities, and is involved in nucleotide metabolism. CD38 uses NAD + It is used as a substrate to form nucleotide metabolites such as cyclic ADP ribose (cADPR), which is an effective second messenger that regulates Ca2+ in the cytoplasm. 2+ Mobilization of CD38 activates signaling pathways that control various biological processes. Studies have also found that CD38 can degrade ATP and NAD + , cADPR, and AMP, thereby producing adenosine, inducing an inhibitory immune microenvironment. In summary, CD38 activity is essential for maintaining NAD + The dynamic balance of substances such as vitamin C and niacinamide is very important.

[0004] Based on the analysis of TCGA data, CD38 is highly expressed in some non-solid and solid tumors, the former including chronic B-lymphocytic leukemia, multiple myeloma, mantle cell lymphoma, etc., and the latter including lung cancer, gastric cancer, melanoma, glioma, esophageal cancer, cervical cancer, etc. Although CD38 is expressed in a variety of human tissues, its expression level in other normal organs is extremely low or low. Moreover, after the primary cancer with high expression of CD38 metastases, the tumor tissue in the metastatic lesion still maintains a high level of CD38 expression. Many studies have shown that CD38 has great potential as a new therapeutic target for malignant tumors and their metastases, and is a very promising target protein molecule.

[0005] At present, some research progress has been made on antibodies targeting CD38. Currently, monoclonal antibodies targeting CD38 that have been used in clinical treatment include daratumumab, isatuximab and MOR202. Researchers have analyzed the co-crystal structure of these antibodies and CD38; constructed fluorescent antibodies based on these antibodies, and developed a method for rapid quantification of cell surface CD38 using flow cytometry; in addition, immunotoxins targeting CD38 have been constructed, showing a highly selective killing effect on multiple myeloma (MM) cells, EC 50 It reaches the pM level, has good efficacy and high specificity and safety.

[0006] Nanobodies, also known as single-domain antibodies, are the smallest antigen-binding fragments obtained from heavy-chain antibodies naturally present in the blood of camelids. Compared with traditional antibodies, nanobodies have the advantages of smaller size (about 15kD), good stability and solubility, easy modification, low production cost, and the ability to recognize hidden or uncommon antigenic sites. Although nanobodies for CD38 have made some progress in recent years, new targeted antibodies or their combinations are still needed to make them more selective, more efficient, less toxic and more convenient for patients to continue to improve the clinical outcomes of patients with related tumors. Summary of the invention

[0007] The present invention provides a nanobody targeting CD38, wherein the nanobody targeting CD38 is any one of the following two antibodies:

[0008] primary antibody and secondary antibody;

[0009] The first antibody and the second antibody each include three complementarity determining regions;

[0010] Wherein, the sequence of CDR1 of the complementary determining region of the first antibody is shown as SEQ.ID NO.1, the sequence of CDR2 is shown as SEQ.ID NO.3, and the sequence of CDR3 is shown as SEQ.ID NO.5;

[0011] The sequence of CDR1 of the complementary determining region of the second antibody is shown in SEQ.ID NO.2, the sequence of CDR2 is shown in SEQ.ID NO.4, and the sequence of CDR3 is shown in SEQ.ID NO.6.

[0012] Preferably, the amino acid sequence of the first antibody is shown as SEQ.ID NO.7.

[0013] Preferably, the amino acid sequence of the second antibody is shown as SEQ.ID NO.8.

[0014] In addition, the present invention also provides a pharmaceutical composition, which contains the CD38-targeting nanobody or its antigen-binding fragment as described above.

[0015] In addition, the present invention also provides a use of a nano-antibody targeting CD38 as described above in the preparation of a drug for diagnosing and / or treating a tumor, wherein the tumor includes one or more of epidermal tissue tumors, connective tissue tumors, lymphatic and hematopoietic system tumors, organ epithelial cell tumors, reproductive system and urinary system tumors, digestive system tumors, nervous system tumors, and endocrine system tumors.

[0016] Preferably, the tumor is a tumor of the respiratory system among epithelial cell tumors of the organ.

[0017] Preferably, the tumor is non-small cell lung cancer among the respiratory system tumors.

[0018] In addition, the present invention also provides the use of the nano-antibody targeting CD38 as described above in the preparation of a product for detecting tumors.

[0019] Preferably, the product for detecting tumors includes at least one of a detection reagent, a detection kit and a detection device.

[0020] The present invention provides a nano antibody targeting CD38, a pharmaceutical composition and its application, wherein the nano antibody targeting CD38 is any one of the following two antibodies: a first antibody and a second antibody; the first antibody and the second antibody both include three complementary determining regions; wherein the sequence of CDR1 of the complementary determining region of the first antibody is shown as SEQ.ID NO.1, the sequence of CDR2 is shown as SEQ.ID NO.3, and the sequence of CDR3 is shown as SEQ.ID NO.5; the sequence of CDR1 of the complementary determining region of the second antibody is shown as SEQ.ID NO.2, the sequence of CDR2 is shown as SEQ.ID NO.4, and the sequence of CDR3 is shown as SEQ.ID NO.6. The present invention obtains two nano antibodies with high affinity to CD38 protein based on phage nano antibody library screening, and verifies their binding to CD38 protein. The two nano antibodies targeting CD38 provided in the present invention have high affinity to purified and cell surface CD38 protein. The specific treatment and / or delivery strategy developed based on these two nanoantibodies (including derivative antibodies modified by adding, reducing or replacing the original sequences of these two nanoantibodies) provides an effective new method for anti-tumor intervention targeting CD38. At the same time, molecular probes and other detection reagents can be prepared based on the nanoantibodies, which provide a new way for tumor-specific imaging and diagnosis by specifically binding to CD38 on the tumor surface. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 The immunofluorescence staining test result of CD38 protein on PC9 cells in the first embodiment of the present invention;

[0022] Figure 2 The immunofluorescence staining results of CD38 protein on A549 cells in Example 1 of the present invention are as follows;

[0023] Figure 3 The Western Blot test results of CD38 expression on PC9 and A549 cells in Example 1 of the present invention;

[0024] Figure 4 The Coomassie Brilliant Blue staining result of the nanobody (fused with His tag and HA tag) targeting CD38 expressed and purified in Example 2 of the present invention (size is about 16kD);

[0025] Figure 5 The Western Blot test results of the nanobody targeting CD38 in the second embodiment of the present invention (using HA and His tag antibodies);

[0026] Figure 6The results of the binding activity between the CD38-targeting nanobody and the CD38 protein detected by ELISA in the second embodiment of the present invention (the results all show binding activity);

[0027] Figure 7 The affinity test result of the first antibody NbD5 in the CD38-targeting nanobody based on the Biacore SPR platform in the second embodiment of the present invention and the CD38 protein (38.26 nM);

[0028] Figure 8 The affinity test result of the second antibody NbD6 in the CD38-targeting nanobody based on the Biacore SPR platform in the second embodiment of the present invention and the CD38 protein (21.07 nM);

[0029] Fig. 9 The results of localization of the CD38-targeting nanobody in PC9 and A549 cells based on laser confocal microscopy in the second embodiment of the present invention (the nanobody can be localized to CD38 on the cell surface);

[0030] Fig.10 The results of the binding of different concentrations of CD38-targeting nanobodies to CD38 antigen on PC9 cells in Example 2 of the present invention;

[0031] Fig.11 The results of the binding of different concentrations of CD38-targeting nanobodies to the CD38 antigen on A549 cells in Example 2 of the present invention.

[0032] The realization of the purpose, functional features and advantages of the present invention will be further explained in conjunction with embodiments and with reference to the accompanying drawings. DETAILED DESCRIPTION

[0033] In an embodiment of the present application, a nanobody targeting CD38 is provided, wherein the nanobody targeting CD38 is any one of the following two antibodies:

[0034] primary antibody and secondary antibody;

[0035] The first antibody and the second antibody each include three complementarity determining regions;

[0036] Wherein, the sequence of CDR1 of the complementary determining region of the first antibody is shown as SEQ.ID NO.1, the sequence of CDR2 is shown as SEQ.ID NO.3, and the sequence of CDR3 is shown as SEQ.ID NO.5;

[0037] The sequence of CDR1 of the complementary determining region of the second antibody is shown in SEQ.ID NO.2, the sequence of CDR2 is shown in SEQ.ID NO.4, and the sequence of CDR3 is shown in SEQ.ID NO.6.

[0038] As mentioned above, the complementary determining region (CDR) of the amino acid sequence is the most frequently mutated and diverse part of the antibody molecule. CDR is located in the junction area of ​​the antibody structure and is mainly composed of three heterochains (Hypervariable Region, HVR). The antigen-specific residues that interact with these regions are called variable regions (variable region, V).

[0039] The main function of CDR is to directly react with specific antigen molecules through the most diverse functional regions in the above antibody molecules, thereby realizing the recognition and binding of antigens in the immune process. Therefore, CDR is an important component to ensure the high specificity of antibody molecules.

[0040] Furthermore, the amino acid sequence of the first antibody is shown in SEQ.ID NO.7.

[0041] Furthermore, the amino acid sequence of the second antibody is shown in SEQ.ID NO.8.

[0042] Therefore, the nanoantibody targeting CD38 provided by the present invention includes a first antibody (named NbD5) and a second antibody (named NbD6); both antibodies have three complementary determining regions, namely CDR1, CDR2 and CDR3, and their corresponding sequence numbers are shown in Table 1, and the specific sequences are shown in Table 2.

[0043] Table 1. Correspondence table of the sequence numbers of the complementary determining region sequences and amino acid sequences in the nanobodies targeting CD38

[0044] Nanobodies targeting CD38 CDR1 CDR2 CDR3 Amino acid sequence Primary Antibody SEQ.ID NO.1 SEQ.ID NO.3 SEQ.ID NO.5 SEQ.ID NO.7 Secondary Antibody SEQ.ID NO.2 SEQ.ID NO.4 SEQ.ID NO.6 SEQ.ID NO.8

[0045] Table 2. Complementarity determining region sequences and amino acid sequences of the first antibody and the second antibody in the CD38-targeting nanobody

[0046]

[0047] In addition, in the examples of the present application, a pharmaceutical composition is also provided, which contains the CD38-targeting nanobody or its antigen-binding fragment as described above.

[0048] In addition, the present invention also provides a use of a nano-antibody targeting CD38 as described above in the preparation of a drug for diagnosing and / or treating a tumor, wherein the tumor includes one or more of epidermal tissue tumors, connective tissue tumors, lymphatic and hematopoietic system tumors, organ epithelial cell tumors, reproductive system and urinary system tumors, digestive system tumors, nervous system tumors, and endocrine system tumors.

[0049] Furthermore, the tumor is a respiratory system tumor among the organ epithelial cell tumors.

[0050] Furthermore, the tumor is non-small cell lung cancer among respiratory system tumors.

[0051] In addition, the present invention also provides a use of the nano-antibody targeting CD38 as described above in the preparation of a detection product for detecting tumors.

[0052] Furthermore, the product for detecting tumors includes at least one of a detection reagent, a detection kit and a detection device.

[0053] As mentioned above, the detection reagent can be a reagent for detecting a specific biomarker or chemical component, and can be designed to combine with a nanobody to improve the specificity and sensitivity of the detection.

[0054] As mentioned above, a test kit may contain all the components required to perform a test, for example, may include but are not limited to reagents, buffers, standards and sometimes test consumables. These kits are intended to simplify the test process and make it more user-friendly.

[0055] The above-mentioned detection equipment can be a device for reading and analyzing the results of the detection reagent or kit, or a device that includes the aforementioned nano-antibody to detect tumors, or plays a role with the detection reagent or detection kit. These devices can be manual or automated, and can include software for data processing and result interpretation.

[0056] Example 1: Verification of CD38 protein expression

[0057] 1. Immunofluorescence staining

[0058] A549 and PC9 cells were revived and cultured for 3 times before being seeded in 24-well plates with slides for subsequent experiments. The specific steps are as follows:

[0059] (1) Place cells on ice, wash three times with PBS, and fix with 4% paraformaldehyde for 10 min;

[0060] (2) Washed three times with PBS and blocked with 3% BSA for 1 hour;

[0061] (3) Dilute CD38 antibody at 1:300 in blocking buffer and incubate with cells at 4°C overnight;

[0062] (4) Wash three times with pre-cooled PBS, dilute Alexa 488-conjugated secondary antibody at 1:1000 in blocking solution, and incubate with cells in the dark for 1 hour;

[0063] (5) Wash the sections three times with pre-cooled PBS, seal the sections with DAPI-containing sealing medium, and detect the expression of CD38 protein using a confocal microscope.

[0064] 2. Western Blot

[0065] A549 and PC9 cells were revived and cultured for 3 times before being seeded in 100 mm culture dishes for subsequent experiments. The specific steps are as follows:

[0066] (1) Digest with trypsin for 3-4 minutes, add 4-5 mL PBS to dilute the trypsin to terminate digestion, and collect the cells after centrifugation;

[0067] (2) Add 200 μL of Lysis Buffer (RIPA + 1× Protease inhibitor) to lyse A549 and PC9 cells;

[0068] (3) Ultrasonication for 30 seconds (energy 25%, working 3 seconds, interval 7 seconds), centrifugation at 14000 rpm / 20 min / 4°C, aspirate 90% of the supernatant (180 μL) into a new EP tube, and measure the protein concentration by BCA method (prepare BCA protein concentration determination at the same time, take new ordinary EP tubes equal to the number of samples, add 54 μL PBS + 6 μL sample);

[0069] (4) 12.5% ​​SDS-PAGE run, transfer to membrane (0.45 μm PVDF membrane, 200 mA, 120 min), block with 5% BSA for 1 h, and then incubate with primary antibody overnight;

[0070] (5) Wash with TBST five times, 5 min each time, incubate with secondary antibody for 1 h, wash with TBST five times, 5 min each time, and develop color.

[0071] Experimental results:

[0072] In this example, the expression profile and prognosis of CD38 in multiple cancer types were first analyzed using the TCGA database. This example focused on verifying the expression of CD38 in non-small cell lung cancer A549 cells and PC9 cells. In this example, immunofluorescence staining was used to detect CD38 expression on the surface of PC9 (refer to Figure 1 ), CD38 is expressed on the surface of A549 (reference Figure 2). Western Blot was used for further verification. Figure 3 The results showed that CD38 was significantly expressed in both A549 and PC9 cells.

[0073] Example 2: Screening and verification of CD38 protein extracellular domain nanoantibodies

[0074] 1. Nanoantibody Screening

[0075] The natural alpaca-derived phage display nanoantibody library was screened using the immunotube method. The selected phage display library had a capacity of 2×10 9 The screening steps are as follows:

[0076] (1) The target protein was coated on an immunotube at a concentration of 50 μg / mL and three rounds of enrichment screening were performed;

[0077] (2) The third round of phage eluate was used for plating, and 96 monoclonal clones were randomly selected for ELISA verification. The ELISA reading was 3 times greater than the corresponding BSA reading and the reading was greater than 0.5 as the positive standard;

[0078] (3) The positive monoclones identified by phage ELISA twice were sent to the company for sequencing to confirm the sequence information;

[0079] (4) designing and synthesizing the screened nanoantibodies based on the sequencing information, and expressing and purifying them through Escherichia coli;

[0080] (5) The affinity of the nanobodies was preliminarily identified using an ELISA affinity test, and the nanobodies with better affinity were selected. After expression and purification, the affinity constants were determined using surface plasmon resonance (SPR).

[0081] 2. Purification and expression of nanobodies

[0082] The nanobody gene sequence was cloned into the pCold-II vector, with a 6×His tag fused to its C-terminus, and a hemagglutinin HA tag was fused and expressed for subsequent detection. The expression and purification steps are as follows:

[0083] (1) To prevent the formation of inclusion bodies and protein degradation, 0.2 mM IPTG was used for induction at 16°C;

[0084] (2) Induce expression in large quantities according to the pre-experimental induction conditions, and break the bacteria at 1300W using a high-pressure sterilizer;

[0085] (3) Centrifuge at 12000g for 45 min at 4°C, and incubate the supernatant with Ni-NTA filler at 4°C for 1 h;

[0086] (4) After purification by Ni-NTA column, different imidazole gradients were used for elution and the eluate was collected, each gradient was about 5 mL. The purity was determined to be >95% by SDS-PAGE. After concentration to a volume of about 3 mL, the concentration of the nanoantibody NbD5 was 5.4 mg / mL and the concentration of NbD6 was 3.97 mg / mL by BCA quantitative measurement.

[0087] 3. ELISA experiment of nanoantibodies

[0088] The ELISA plate was coated with CD38 protein overnight and blocked, and then various concentrations of HA-tagged nanoantibodies were added and incubated at room temperature for 1 hour, rinsed 3 times with PBST, and incubated with anti-HA antibody at room temperature for 1 hour. The signal was amplified by horseradish peroxidase (HRP)-labeled anti-HA antibody, and TMB was used for color development. At the same time, controls with irrelevant nanoantibodies and blank controls with irrelevant protein antigens were performed.

[0089] 4. Surface plasmon resonance experiment

[0090] This experiment is used to verify the direct interaction between the purified nanobodies expressed in vitro and the purified antigen protein in vitro, and to calculate the equilibrium constant between the two. The purified antigen protein (CD38) was fixed on the chip, and different concentrations of nanobodies (NbD5 and NbD6) were added sequentially to analyze the affinity with the antigen protein, and the reaction signal within 300s was recorded, the kinetic curve was made, and the relevant parameters were calculated.

[0091] 5. Targeted detection of cells by nanoantibodies

[0092] PC9 and A549 cells were revived and cultured for 3 times before being seeded in 96-well plates for subsequent experiments. The specific steps are as follows:

[0093] (1) Place cells on ice, wash three times with PBS, and fix with 4% paraformaldehyde for 10 min;

[0094] (2) Wash with PBS three times and block with 5% BSA for 1 h;

[0095] (3) The nanoantibodies were diluted with 1% BSA to different concentration gradients (0-1000 nM) and incubated with the cells for 1 h;

[0096] (4) Wash three times with pre-cooled PBST, dilute anti-HA antibody at 1:1000 with 1% BSA, and incubate with cells for 1 h;

[0097] (5) Wash three times with pre-cooled PBST, dilute Alexa 488-conjugated secondary antibody at 1:1000 with 1% BSA, and incubate with cells in the dark for 1 h;

[0098] (6) Wash three times with pre-cooled PBST and detect the binding of nanoantibodies on the cell surface using an automatic electrophoresis fluorescence immunoassay analyzer.

[0099] Experimental results:

[0100] (1) First, the phage nanoantibody library was screened against CD38, and two positive clones were preliminarily identified, which were named NbD5 (first antibody) and NbD6 (second antibody) (the specific sequences and complementary determining regions (CDRs) of NbD5 and NbD6 are shown in Tables 1 and 2).

[0101] (2) Expression and purification of nanobodies (first antibody NbD5 and second antibody NbD6) and control nanobody C9, detection and confirmation of the correctness of the antibody proteins ( Figure 4 and Figure 5 ).Depend on Figure 4 It can be seen that the Coomassie Brilliant Blue staining results showed that by expressing and purifying the nanobody protein fused with the His tag and the HA tag, high-purity nanobodies (NbD5 and NbD6) were obtained, with a size of about 16 kD; Figure 5 It can be seen that the Western Blot detection using HA and His tag antibodies confirmed the correct expression of the nanobody protein.

[0102] (3) Verified by ELISA, such as Figure 6 As shown, it was confirmed that nanobodies NbD5 (first antibody) and NbD6 (second antibody) had binding activity with recombinant CD38 protein.

[0103] (4) The affinity constants of NbD5 and NbD6 clones to recombinant CD38 protein were detected and determined using the Biacore SPR platform. The results showed that the affinity of NbD5 (primary antibody) and NbD6 (secondary antibody) to CD38 protein was 38.26 nM, respectively (ref. Figure 7 ) and 21.07nM (reference Figure 8 ).

[0104] (5) Immunofluorescence and Cell ELISA were used to detect whether the nanoantibody can bind to the CD38 protein on the cell membrane surface. Fig. 9 Under laser confocal microscopy, nanoantibodies NbD5 (first antibody, concentration of 1 μM) and NbD6 (second antibody, concentration of 1 μM) can be localized on CD38 on the surface of PC9 cells and A549 cells, where the left column is the first antibody NbD5 and the right column is the second antibody NbD6; Fig.10 Cell ELISA was used to detect the binding of different concentrations of the first antibody NbD5, the second antibody NbD6 and the control nanobody C9 to the CD38 antigen on PC9 cells; Fig.11 Cell ELISA was used to detect the binding of different concentrations of the first antibody NbD5, the second antibody NbD6 and the control nanobody C9 to the CD38 antigen on A549 cells. The results showed that the nanobodies NbD5 and NbD6 had specific recognition ability for the CD38 protein on the surface of PC9 and A549 cell membranes.

[0105] In summary, the two CD38-targeting nanobodies provided in the present invention have high affinity for both purified and cell-surface CD38 proteins. The specific treatment and / or delivery strategies developed based on these two nanobodies (including derivative antibodies modified by adding, reducing or replacing the original sequences of these two nanobodies) provide an effective new method for anti-tumor intervention targeting CD38. At the same time, molecular probes and other detection reagents can be prepared based on the nanobodies, which specifically bind to CD38 on the surface of tumors, providing a new approach for tumor-specific imaging and diagnosis.

[0106] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present application. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, without contradiction.

[0107] Although the embodiments of the present application have been shown and described above, it can be understood that the above embodiments are exemplary and cannot be understood as limitations on the present application. Ordinary technicians in this field can change, modify, replace and modify the above embodiments within the scope of the present application.

Claims

1. A nanobody targeting CD38, characterized in that: The nanobody targeting CD38 is any one of the following two antibodies: primary antibody and secondary antibody; The first antibody and the second antibody each include three complementarity determining regions; Wherein, the sequence of CDR1 of the complementary determining region of the first antibody is shown as SEQ.ID NO.1, the sequence of CDR2 is shown as SEQ.ID NO.3, and the sequence of CDR3 is shown as SEQ.ID NO.5; The sequence of CDR1 of the complementary determining region of the second antibody is shown in SEQ.ID NO.2, the sequence of CDR2 is shown in SEQ.ID NO.4, and the sequence of CDR3 is shown in SEQ.ID NO.

6.

2. The nanobody targeting CD38 according to claim 1, characterized in that The amino acid sequence of the first antibody is shown in SEQ.ID NO.

7.

3. The nanobody targeting CD38 according to claim 1, characterized in that: The amino acid sequence of the second antibody is shown in SEQ.ID NO.

8.

4. A pharmaceutical composition, characterized in that It contains the CD38-targeting Nanobody or its antigen-binding fragment as described in any one of claims 1 to 3.

5. Use of a nanobody targeting CD38 as claimed in any one of claims 1 to 3 in the preparation of a drug for diagnosing and / or treating tumors, characterized in that: The tumor includes one or more of epidermal tissue tumors, connective tissue tumors, lymphatic and hematopoietic system tumors, organ epithelial cell tumors, reproductive system and urinary system tumors, digestive system tumors, nervous system tumors, and endocrine system tumors.

6. The use according to claim 5, characterized in that The tumor is a respiratory system tumor among epithelial cell tumors of the organ.

7. The use according to claim 5, characterized in that The tumor is non-small cell lung cancer among respiratory system tumors.

8. Use of the CD38-targeting nanobody according to any one of claims 1 to 3 in the preparation of a product for detecting tumors.

9. The use according to claim 8, characterized in that The product for detecting tumors includes at least one of a detection reagent, a detection kit and a detection device.

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