Use of CD38 inhibitor for blocking mononuclear macrophage function in treating itp

Abstract

Use of a CD38 inhibitor for blocking mononuclear macrophage function in treating ITP. An anti-CD38 antibody serving as a CD38 inhibitor specifically binds to CD38 molecules on the membrane surface of mononuclear macrophages to inhibit the proliferation of mononuclear macrophages, reducing the number and proportion of mononuclear macrophages, effectively inhibiting the ADCC effect, and rapidly improving platelet counts in patients. The inhibitor can be used for treating various types of ITP patients, and the effective rate in treating ITP is high.

Description

Use of a CD38 inhibitor blocking monocyte macrophage function in the treatment of ITP TECHNICAL FIELD

[0001] The present application relates to the field of biological medicine, and particularly relates to use of a CD38 inhibitor blocking monocyte function in treatment of ITP. BACKGROUND

[0002] Primary immune thrombocytopenia (ITP) is an organ-specific thrombocytopenia characterized by isolated peripheral blood platelet reduction (<100×10 9 / L). The annual incidence of adult ITP is 9 to 20 per 100,000. The main clinical symptoms of the disease include skin mucosa bleeding and obvious fatigue, and the severity of bleeding is directly related to the number of platelets. Although most ITP patients only show a slight bleeding tendency, when the platelet count drops to 30×10 9 / L or less, some patients may face life-threatening bleeding events such as gastrointestinal bleeding and intracranial hemorrhage. At present, the first-line drugs for ITP treatment include glucocorticoids and gamma globulin. Although the initial response rate of glucocorticoids can reach 70%, the sustained response rate after drug withdrawal is only 20-40%. Given the side effects (such as hypertension, hyperglycemia and obesity) that may be caused by long-term use of glucocorticoids, current guidelines do not recommend it as a sustained treatment regimen for ITP. Gamma globulin can quickly increase the peripheral blood platelet count of patients and is currently mainly used for emergency treatment of ITP or combined with other treatment methods. For patients who do not respond to first-line treatment or relapse, the recommended second-line treatment strategy includes thrombopoietin receptor agonists, rituximab and splenectomy. Although there are currently a variety of treatment options available, the response of ITP patients to these therapies shows significant individual differences. Therefore, there is an urgent need to develop safer treatment strategies to improve the initial response rate and ensure the stability of long-term efficacy, so as to improve the treatment status of ITP.

[0003] The pathophysiology of ITP is complex, and the core is that the body loses immune tolerance to platelet autoantigens and produces anti-platelet autoantibodies. This process triggers abnormal activation of humoral and cellular immunity, leading to increased antibody-mediated platelet destruction and decreased megakaryocyte platelet production capacity. In the pathogenesis of ITP, antibody-dependent cell-mediated cytotoxicity (ADCC) plays a key role, and it is also one of the important mechanisms for the body to clear cells infected by pathogens or expressing abnormal antigens. As the main executors in the ADCC process, natural killer cells and mononuclear macrophages can quickly clear platelets combined with autoantibodies, causing a significant decrease in the number of peripheral platelets in ITP patients. Studies have shown that gamma globulin can reduce ADCC-mediated platelet clearance by promoting the apoptosis of natural killer cells in the spleen of ITP patients. At the same time, the expression of major histocompatibility complex (MHC) molecules on the surface of megakaryocytes and platelets may enable them to play a role similar to antigen-presenting cells to activate T cells, thereby accelerating the self-clearance of platelets. Early studies have shown that specific anti-platelet plasma cells in the spleen of ITP patients can migrate to the bone marrow and survive as long-lived plasma cells (LLPCs), which are the main source of natural antibodies in the body and are also a key factor for the recurrence of ITP patients after immunosuppressive treatment such as rituximab.

[0004] The monoclonal antibody currently used in clinical treatment of ITP is anti-CD20 mAb (rituximab, etc.). CD20 molecules are widely expressed on the surface of pre-B and mature B lymphocytes, and monoclonal antibodies targeting CD20 can clear all CD20-positive B cells in the body, including autoreactive B cells (producing autoantibodies), by initiating antibody-dependent cytotoxicity, complement-dependent cytotoxicity, etc. Anti-CD20 mAb treatment of ITP has a slow onset, with a median response time (platelet count rising to 30 x 10 9 / L or more) of about 4-6 weeks, and patients still have bleeding risk at the early stage of treatment. In addition, the short-term complete response rate (platelet count ≥ 100 x 10 9 / L) of this treatment is about 60%, but the 5-year sustained effective rate is only 20-30%. However, since plasma cells and plasmablasts do not express CD20 on their surface, B cell clearance treatment targeting CD20 cannot completely clear the antibody-secreting cells in the patient's body, and may even stimulate short-lived plasma cells in the spleen to reprogram into long-lived plasma cells, causing reactivation of autoimmune B cell response and leading to disease recurrence.

[0005] Therefore, it is necessary to develop a drug that can provide long-term remission for ITP patients and reduce ITP recurrence.

[0006] SUMMARY

[0007] To solve the above technical problems, the present application provides the use of an anti-CD38 antibody in the preparation of a medicament for treating ITP, and the indication of the anti-CD38 antibody for treating ITP, which mainly includes that the anti-CD38 antibody can rapidly increase the peripheral platelet count of ITP patients and can be used for the emergency treatment of ITP patients.

[0008] In a first aspect of the present application, the use of an anti-CD38 antibody in the preparation of a medicament for treating ITP is provided, and the anti-CD38 antibody can inhibit the proliferation of mononuclear macrophages to increase the peripheral platelet count.

[0009] Preferably, the medicament for treating ITP includes an anti-CD38 antibody.

[0010] Preferably, the anti-CD38 antibody inhibits the expression of serum antibodies.

[0011] Preferably, the serum antibodies include IgG, IgM and / or IgA.

[0012] Preferably, the anti-CD38 antibody reduces the proliferation of CD38 in mononuclear macrophages of ITP patients, reduces the ADCC function mediated by mononuclear macrophages, and reduces the destruction of platelets.

[0013] In a specific embodiment of the present application, the anti-CD38 antibody specifically binds to the CD38 molecules on the membrane surface of mononuclear macrophages, reduces the number and proportion of mononuclear macrophages, reduces the expression of serum antibodies, and further inhibits ADCC to increase the platelet count of ITP patients.

[0014] Preferably, the anti-CD38 antibody is an anti-CD38 antibody that can block the activity of mononuclear macrophages.

[0015] Preferably, the ITP patient includes a platelet count <30x10 9 / L.

[0016] Preferably, the primary immune thrombocytopenia includes persistent or chronic primary immune thrombocytopenia; wherein the patient with persistent or chronic primary immune thrombocytopenia is a patient whose platelet count has been continuously reduced for more than 3 months after diagnosis.

[0017] More preferably, the primary immune thrombocytopenia includes adult persistent or chronic primary immune thrombocytopenia.

[0018] Preferably, the primary immune thrombocytopenia includes relapsed and / or refractory primary immune thrombocytopenia.

[0019] Preferably, the ITP patients include patients who have received one or more prior treatment.

[0020] Preferably, the prior treatment includes glucocorticoid therapy, immunoglobulin therapy, molecular targeted therapy, thrombopoietic drug therapy or immunosuppressant therapy.

[0021] Preferably, the glucocorticoid includes, but is not limited to, prednisone, methylprednisone, prednisone acetate, prednisolone, methylprednisolone, prednisolone acetate, prednisolone sodium succinate, methylprednisolone sodium succinate, betamethasone, beclometasone dipropionate, hydrocortisone or dexamethasone.

[0022] Preferably, the immunoglobulin includes immunoglobulin for injection. In some embodiments of the present application, the immunoglobulin can be gamma globulin.

[0023] Preferably, the drug for molecular targeted therapy includes, but is not limited to, rituximab, veltuzumab, dalizumab, basiliximab, alemtuzumab or ocrelizumab.

[0024] Preferably, the thrombopoietic drug includes, but is not limited to, eltrombopag, avatrombopag, atrombopag, romiplostim, lusombrug or recombinant human thrombopoietin (rhTPO).

[0025] Preferably, the other drugs that can be used for the treatment of immune thrombocytopenia include, but are not limited to, immunosuppressants, such as sodium ganciclovir, imatinib, cyclosporin (A), mycophenolate sodium, mycophenolate mofetil (mycophenolate), danazol, azathioprine, psoralen, methotrexate, hydroxychloroquine, amphenicol, cyclophosphamide, thalidomide, azithromycin, montelukast, sorafenib, lucatumumab, decitabine, vincristine, alefacept, tretinoin.

[0026] Preferably, the ITP patients include patients who have received one or more prior treatment.

[0027] Preferably, the drug for treating ITP also includes methylprednisolone, acetaminophen, antihistamines and / or drugs for the prevention of herpes zoster.

[0028] Preferably, the antihistamines include, but are not limited to, chlorpheniramine, promethazine, loratadine or montelukast sodium tablets.

[0029] The drugs for the prevention of herpes zoster include, but are not limited to, acyclovir tablets, valacyclovir hydrochloride tablets or famciclovir tablets.

[0030] Preferably, the administration of the drug includes, but is not limited to, oral administration, enteral administration, subcutaneous injection, intramuscular injection, intravenous injection, nasal administration, transdermal administration, subconjunctival administration, intraocular administration, orbit administration, retrobulbar administration, retinal administration, choroidal administration, intraperitoneal injection or intrathecal injection, etc.

[0031] Preferably, the anti-CD38 antibody is administered at a concentration of 10-20 (10, 12, 14, 15, 16, 18, 20) mg / kg, and more preferably 16 mg / kg.

[0032] Preferably, the anti-CD38 antibody is administered at least once, and the administration frequency is once a week.

[0033] In a specific embodiment, the anti-CD38 antibody is administered at a concentration of 16 mg / kg by intravenous infusion once a week for 8 weeks of continuous treatment.

[0034] Preferably, the drug for treating ITP other than the anti-CD38 antibody is administered before, after or simultaneously with the administration of the anti-CD38 antibody.

[0035] Preferably, methylprednisolone, acetaminophen and / or antihistamines are administered before the administration of the anti-CD38 antibody.

[0036] Preferably, methylprednisolone and / or zoster prophylaxis are administered after the administration of the anti-CD38 antibody.

[0037] The drug dosage form includes powder, tablet, sustained-release tablet, chewable tablet, effervescent tablet, lozenge, buccal tablet, sublingual tablet, capsule, fine granule, granule, such as oral preparations, such as pills, dry syrup, solution, suspension, syrup and elixir, as well as eye drops, eye drops, eye ointment, injection, infusion, external preparation, etc.

[0038] Preferably, the medicine further comprises pharmaceutically acceptable excipients and other pharmaceutically acceptable known pharmaceutical additives of the dosage form, which can be appropriately mixed into the pharmaceutical composition according to its physicochemical properties, biological properties, etc. For example, excipients (lactose, starch, crystalline cellulose, sodium phosphate, etc.), solvents (water, soybean oil, physiological saline, non-aqueous solvent for injection, etc.), binders (starch, gelatin, acacia, sodium alginate, sodium carmellose, methyl cellulose, ethyl cellulose, hydroxypropyl methyl cellulose, hydroxypropyl cellulose, polyvinyl alcohol pyrrolidone, etc.), disintegrants (starch, carboxymethyl cellulose sodium, etc.), lubricants (talc, magnesium stearate, calcium stearate, polyethylene glycol, sucrose fatty acid ester, etc.), coating agents (sucrose, hydroxypropyl cellulose (HPC), shellac, gelatin, glycerol, hydroxypropyl methyl cellulose, hydroxypropyl methyl cellulose phthalate, cellulose acetate phthalate, etc.), stabilizers (sodium bisulfite, sodium thiosulfate, sodium edetate, sodium citrate, ascorbic acid, dibutyl hydroxytoluene, etc.), preservatives (methyl paraben, ethyl paraben, propyl paraben, benzyl alcohol, phenol, chlorobutanol, benzalkonium chloride, benzethonium chloride, sodium dehydroacetate, thiomersal, etc.), thickening agents (methyl cellulose, carboxymethyl cellulose sodium, chondroitin sulfate sodium, sodium alginate, etc.), suspending agents (various non-ionic surfactants, methyl cellulose, carboxymethyl cellulose sodium, etc.), emulsifiers (acacia, cholesterol, sorbitan sesquioleate, polysorbate 80, sodium dodecyl sulfate, etc.), buffers (citric acid, acetic acid, sodium phosphate, boric acid), surfactants (hydrogenated castor oil, polysorbate 80, etc.), coloring agents (water-soluble food colorants, lake pigments, etc.), flavoring agents (lactose, sucrose, glucose, mannitol, etc.), flavoring agents (aromatic essential oils, etc.), plasticizers (phthalate esters, vegetable oils, polyethylene glycol, etc.).

[0039] In a second aspect of the present application, an anti-CD38 antibody is provided for use in inhibiting monocyte macrophage proliferation.

[0040] Preferably, the anti-CD38 antibody inhibits monocyte macrophage proliferation to increase peripheral platelet count.

[0041] Preferably, the anti-CD38 antibody inhibits serum antibody expression.

[0042] Preferably, the serum antibody comprises IgG, IgM, and / or IgA.

[0043] Preferably, the anti-CD38 antibody inhibits CD38 expression in monocyte macrophages, reduces monocyte macrophage-mediated ADCC function, and reduces platelet destruction.

[0044] In one embodiment of the present application, the anti-CD38 antibody reduces the number and proportion of mononuclear macrophages by specifically binding to CD38 molecules on the membrane surface of mononuclear macrophages, reduces serum antibody expression, and thereby inhibits ADCC, thereby increasing the platelet count in ITP patients.

[0045] Preferably, the anti-CD38 antibody is an anti-CD38 antibody that can block the activity of mononuclear macrophages.

[0046] In a third aspect of the present application, a pharmaceutical composition comprising an anti-CD38 antibody and a pharmaceutically acceptable excipient is provided.

[0047] Preferably, the anti-CD38 antibody is administered at a concentration of 10-20 (10, 12, 14, 15, 16, 18, 20) mg / kg, and more preferably at a concentration of 16 mg / kg.

[0048] Preferably, the pharmaceutical composition further comprises methylprednisolone, acetaminophen, an antihistamine, and / or a zoster prophylactic.

[0049] Preferably, the antihistamine includes, but is not limited to, chlorpheniramine, promethazine, loratadine, or montelukast sodium tablets.

[0050] The zoster prophylactic includes, but is not limited to, acyclovir tablets, valacyclovir hydrochloride tablets, or famciclovir tablets.

[0051] Preferably, the pharmaceutical composition is administered by a method including, but not limited to, oral administration, enteral administration, subcutaneous injection, intramuscular injection, intravenous injection, nasal administration, transdermal administration, subconjunctival administration, intraocular administration, orbital administration, retrobulbar administration, retinal administration, choroidal administration, intraperitoneal injection, intrathecal injection, and the like.

[0052] Preferably, the anti-CD38 antibody is administered at least once, with a frequency of once a week.

[0053] In one embodiment, the anti-CD38 antibody is administered at a dose of 16 mg / kg by intravenous infusion once a week for 8 weeks of continuous treatment.

[0054] The pharmaceutical dosage form includes a powder, a tablet, a sustained-release tablet, a chewable tablet, an effervescent tablet, a lozenge, a buccal tablet, a sublingual tablet, a capsule, a fine granule, a granule, for example, an oral preparation such as a pill, a dry syrup, a solution, a suspension, a syrup, and an elixir, as well as an eye drop, an eye drop, an eye ointment, an injection, an infusion, an external preparation, and the like.

[0055] Preferably, the medicine further comprises pharmaceutically acceptable excipients and other pharmaceutically acceptable known pharmaceutical additives of the dosage form, which can be appropriately mixed into the pharmaceutical composition according to its physicochemical properties, biological properties, etc. For example, excipients (lactose, starch, crystalline cellulose, sodium phosphate, etc.), solvents (water, soybean oil, physiological saline, non-aqueous solvent for injection, etc.), binders (starch, gelatin, acacia, sodium alginate, sodium carmellose, methyl cellulose, ethyl cellulose, hydroxypropyl methyl cellulose, hydroxypropyl cellulose, polyvinyl alcohol pyrrolidone, etc.), disintegrants (starch, carboxymethyl cellulose sodium, etc.), lubricants (talc, magnesium stearate, calcium stearate, polyethylene glycol, sucrose fatty acid ester, etc.), coating agents (sucrose, hydroxypropyl cellulose (HPC), shellac, gelatin, glycerol, hydroxypropyl methyl cellulose, hydroxypropyl methyl cellulose phthalate, cellulose acetate phthalate, etc.), stabilizers (sodium bisulfite, sodium thiosulfate, sodium edetate, sodium citrate, ascorbic acid, dibutyl hydroxytoluene, etc.), preservatives (methyl paraben, ethyl paraben, propyl paraben, benzyl alcohol, phenol, chlorobutanol, benzalkonium chloride, benzethonium chloride, sodium dehydroacetate, thiomersal, etc.), thickening agents (methyl cellulose, carboxymethyl cellulose sodium, chondroitin sulfate sodium, sodium alginate, etc.), suspending agents (various non-ionic surfactants, methyl cellulose, carboxymethyl cellulose sodium, etc.), emulsifiers (acacia, cholesterol, sorbitan sesquioleate, polysorbate 80, sodium lauryl sulfate, etc.), buffers (citric acid, acetic acid, sodium phosphate, boric acid), surfactants (hydrogenated castor oil, polysorbate 80, etc.), coloring agents (water-soluble food dyes, lake pigments, etc.), flavoring agents (lactose, sucrose, glucose, mannitol, etc.), flavoring agents (aromatic essential oils, etc.), plasticizers (phthalate esters, vegetable oils, polyethylene glycol, etc.).

[0056] The "application" in the present application does not involve the diagnosis or treatment of diseases.

[0057] The subject / patient of the medicine in the present application can be a mammal, such as a human, a monkey, a dog, a rabbit, a mouse, a rat, etc.

[0058] The term "comprising" or "including" in the present application is an open description containing the specified components or steps described, and other specified components or steps that do not materially affect.

[0059] The beneficial effects of the present application:

[0060] CD38 molecule is highly expressed on the surface of plasma cells, including long-lived plasma cells. Targeting CD38 for cell clearance therapy can effectively eliminate antibody-secreting cells in the bone marrow of ITP patients, providing the possibility of long-term remission for ITP patients. More importantly, CD38 molecules also exist widely on the surface of mononuclear macrophages, effectively inhibiting antibody-dependent cell-mediated cytotoxicity (ADCC), rapidly increasing the platelet count of patients (the median platelet count of patients can reach more than 100x109 / L after 1 week of treatment), and significantly reducing the risk of bleeding in patients. The clinical effect of this scheme is significant, and the effect is durable and stable, which can be applied to the treatment of various types of thrombocytopenia patients.

[0061] The present study closely monitors the dynamic changes of platelet count in patients before and after receiving anti-CD38 treatment, and finds that anti-CD38 antibody can rapidly increase the peripheral blood platelet count of ITP patients and can be used for emergency treatment of ITP patients. The efficiency of anti-CD38 antibody in treating ITP is high, and the short-term response rate is about 96% (the proportion of platelet count ≥50x109 / L during 8 weeks of drug use), and the 24-week efficiency is about 64%. And the present application first proposes and proves that anti-CD38 antibody can reduce the platelet destruction ability of mononuclear macrophages by blocking CD38 molecules on the surface of mononuclear macrophages, rapidly increasing the peripheral blood platelet count of ITP patients. BRIEF DESCRIPTION OF DRAWINGS

[0062] Figure 1 is a platelet count fold line chart of patients receiving anti-CD38 monoclonal antibody treatment.

[0063] Figure 2 is a modeling strategy of ITP passive mouse model.

[0064] Figure 3 is the flow cytometry detection result of human peripheral blood, A is the distribution proportion of CD38 expressing cells, B is the average fluorescence intensity of CD38 expression, *: p<0.05, **: p≤0.01, ***: p≤0.001, HC is healthy control, P-d0 is ITP patient.

[0065] Figure 4 is the expression level of CD38 on the surface of each immune cell in peripheral blood, A is the detection result of CD14+ mononuclear cells, B is the IgG level in serum, C is the IgA level in serum, D is the IgM level in serum, ***: p≤0.001.

[0066] Figure 5 is the platelet count monitoring result of tail vein blood routine after treating ITP mice with anti-mouse CD38 monoclonal antibody.

[0067] Figure 6 is the detection result of spleen mononuclear cells of mice after anti-mouse CD38 monoclonal antibody treatment, wherein A is the number of mononuclear cells, B is the proportion of Ly6C high expression mononuclear cells expressing CD38, C is the proportion of Ly6C low expression mononuclear cells expressing CD38, **: p≤0.01.

[0068] Figure 7: Detection results of mouse spleen macrophages after anti-mouse CD38 monoclonal antibody treatment, wherein A is the number of macrophages, B is the number of CD38-expressing macrophages, C is the proportion of CD38-expressing macrophages, **: p≤0.01. DETAILED DESCRIPTION

[0069] In order to enable persons skilled in the art to better understand the technical solutions in the present application, the technical solutions in the embodiments of the present application will be clearly and completely described below. Obviously, the described embodiments are only some of the embodiments of the present application, not all. Based on the embodiments in the present application, all other embodiments obtained by persons skilled in the art without creative labor should fall within the scope of protection of the present application.

[0070] The materials, reagents, instruments, etc. used in the following examples can be obtained from commercial channels unless otherwise specified.

[0071] Example 1 Clinical efficacy of anti-CD38 monoclonal antibody for treating ITP

[0072] Implementation details:

[0073] Patient population: Adult ITP patients (age ≥18 years old) diagnosed with primary immune thrombocytopenia with persistence or chronicity (course of disease more than 3 months).

[0074] Main inclusion criteria:

[0075] ① Failure of previous glucocorticoid therapy (ineffective, or unable to maintain efficacy or dependent, or relapse); and failure of at least one guideline-recommended second-line treatment for ITP (including rituximab and / or thrombopoietin drugs such as rhTPO or eltrombopag, etc.) (ineffective, or unable to maintain efficacy, or relapse), ineffective defined as: rhTPO 300U / kg / d >14 days, eltrombopag 75mg / d >28 days, avatrombopag 7.5mg / d >28 days, aravatrombopag 60mg / d >28 days, platelet count still <30×10 9 / L.

[0076] ② Platelet count <30×10 9 / L within 48 hours before the first administration of the study drug (at least two consecutive platelet counts less than 30×10 9 / L (two tests at least 1 day apart) during the screening visit and / or before administration of the study drug).

[0077] ③ Allow inclusion of subjects on stable dose maintenance therapy (including concomitant medications including glucocorticoids (≤0.5 mg / kg prednisone or equivalent) or TPO receptor agonists, etc., but limited to only one stable dose concomitant medication at the time of enrollment, which must have been on a stable dose for at least 4 weeks prior to first dose).

[0078] ④ For female patients of childbearing potential, a negative pregnancy test result is required. Female patients of childbearing potential and male patients must take highly effective contraceptive measures during the study period and for 4 months / 6 months after treatment, respectively.

[0079] ⑤ ECOG performance status score ≤2.

[0080] Major exclusion criteria:

[0081] ① Allergic to CD38 mAb or excipients, or who have previously received CD38 mAb and were ineffective.

[0082] ② Associated with autoimmune hemolytic anemia, or various secondary and hereditary thrombocytopenia.

[0083] ③ History of any thrombotic or embolic events or associated with extensive and severe bleeding within 12 months prior to first dose; or use of anticoagulants or any drugs with anti-platelet effects within 3 weeks prior to first dose.

[0084] ④ Received emergency treatment for ITP (e.g., methylprednisolone, platelet, gamma globulin infusion or TPO receptor agonist treatment) within 2 weeks prior to first dose.

[0085] ⑤ Received splenectomy within 6 months prior to first dose; previously received allogeneic stem cell transplantation or organ transplantation; history of severe recurrent or chronic infections.

[0086] ⑥ Showed clinically significant laboratory abnormalities at screening:

[0087] a) Alanine aminotransferase or aspartate aminotransferase ≥ upper limit of normal (ULN).

[0088] b) Total bilirubin ≥ 1.2 times ULN (Note: Patients with documented Gilbert's syndrome recorded in the patient's medical history cannot be excluded according to this criterion).

[0089] c) Creatinine, urea nitrogen ≥ ULN.

[0090] ⑦HIV antibody or syphilis antibody positive; screening with hepatitis B surface antigen (HBsAg) positive, or hepatitis B core antibody positive, polymerase chain reaction (PCR) test results for HBV-DNA positive (if hepatitis B core antibody positive but HBV-DNA negative, generally can be included in the group, but need to monitor HBV-DNA every 4 weeks), or hepatitis C virus (HCV) antibody positive, etc.

[0091] Treatment regimen: anti-CD38 monoclonal antibody 16 mg / kg intravenous infusion, once a week, continuous treatment for 8 weeks. During the medication, the vital signs of the patients were closely observed, and the blood routine was regularly detected.

[0092] Pre-medication before and after infusion:

[0093] Before infusion (within 1-2 hours):

[0094] ① Methylprednisolone: 100 mg of methylprednisolone (or equivalent drugs) was given intravenously before the first and second medication, and since the third treatment, the dose of glucocorticoid was reduced to 60 mg of methylprednisolone (or equivalent drugs) before each medication, which was given orally or intravenously.

[0095] ② About 1 hour or less before infusion, acetaminophen 500 mg-1000 mg was taken orally.

[0096] ③ About 1 hour before infusion, diphenhydramine 25-50 mg was taken orally (or other antihistamines with equivalent dose, such as chlorpheniramine 8 mg, taken orally, promethazine 25 mg, taken orally; loratadine 10 mg taken orally, montelukast sodium tablets 10 mg taken orally).

[0097] After infusion:

[0098] On the first and second days after the first and second infusion of anti-CD38 monoclonal antibody, methylprednisolone 20 mg was given orally or intravenously; since the third infusion, no methylprednisolone treatment was given after the end of anti-CD38 monoclonal antibody infusion.

[0099] Zoster prophylaxis: within 1 week after starting CM313 (humanized monoclonal antibody targeting CD38), antiviral prophylaxis (acyclovir 0.2 g, taken orally, twice a day) was started, which lasted for more than 3 months to prevent herpes zoster reactivation.

[0100] Primary outcome:

[0101] Platelet count (x10 9Results are shown in Figure 1 and Table 1. The dynamic changes of platelet count in 21 patients before and after anti-CD38 treatment were closely monitored. Anti-CD38 monoclonal antibody can rapidly increase the peripheral blood platelet count of ITP patients and can be used for emergency treatment of ITP patients. Anti-CD38 monoclonal antibody treatment of ITP has high efficiency, and the short-term response rate is about 96% (platelet count ≥50x109 / L during 8 weeks of drug use), and the 24-week efficiency is about 64%. 9 / L ratio), and the 24-week efficiency is about 64%.

[0102] Table 1 Platelet count (x109 / L) of patients at each visit point after receiving anti-CD38 monoclonal antibody treatment 9 / L

[0103] Example 2: Anti-CD38 monoclonal antibody can reduce platelet destruction by blocking mononuclear macrophage function

[0104] Implementation details:

[0105] 1) Human peripheral blood flow detection:

[0106] Peripheral blood of age and gender matched healthy volunteers and patients receiving anti-CD38 monoclonal antibody treatment at different visit points (0d, 7d, 21d, 105d) was centrifuged to separate plasma and whole blood at room temperature. The separated whole blood (200ul / tube) was transferred to a flow tube, and fluorescently labeled mouse anti-human monoclonal antibodies (CD14, CD3, CD8, CD19, CD38, CD56, etc.) were added at room temperature in the dark for 15 minutes. An appropriate amount of red blood cell lysate was added to the flow tube at the end of incubation, shaken well and incubated for 10 minutes. After washing twice with phosphate buffered saline, the supernatant was discarded after centrifugation, and the cell pellet was resuspended with 200ul 2% tissue cell fixative. Canto II flow cytometer was used for detection, and FlowJo software was used for result analysis.

[0107] 2) Establishment of passive ITP mouse model:

[0108] The modeling strategy of ITP passive mouse model is shown in Figure 2. After tail vein blood sampling of 8-10 week old female BALB / c mice, rat anti-mouse CD41 monoclonal antibody was given intraperitoneally (once a day, -2d dose of 68ug / kg, -1d of 102ug / kg, 0d-+2d of 136ug / kg). The blood routine of mice was monitored daily by tail vein blood sampling.

[0109] 3) Anti-mouse CD38 monoclonal antibody treatment of ITP passive model mice and flow cytometry detection of splenic mononuclear macrophages:

[0110] 8-10 weeks old female BALB / c mice were randomly divided into two groups (control group and treatment group), and both groups of mice were given intraperitoneal injection of anti-CD41 monoclonal antibody to construct a passive ITP model according to the steps described in 2). At the same time, on d0, the treatment group was given intravenous injection of anti-mouse CD38 monoclonal antibody (10 mg / kg, 12 hours apart from anti-CD41 monoclonal antibody), and the control group was given the same volume of normal saline. Daily tail vein blood sampling was continued to monitor blood routine.

[0111] +3d, after weighing, the mice were anesthetized and sacrificed, and the spleen was dissected and removed. After weighing the spleen, a single cell suspension was prepared by grinding on ice. An appropriate amount of prepared single cell suspension was taken in a flow tube, and fluorescently labeled rat anti-mouse monoclonal antibody (CD45, Ly6C, Ly6G, F4 / 80, etc.) was added and mixed, then incubated at 4°C for 30 minutes. After incubation, an appropriate amount of red cell lysate was added to the flow tube, mixed and shaken for 5 minutes, then washed twice with phosphate buffered saline, centrifuged to discard the supernatant, and the cell pellet was resuspended in an appropriate amount of phosphate buffered saline. Canto II flow cytometry was used for detection, and FlowJo software was used for analysis of the results.

[0112] Main results:

[0113] Human peripheral blood data:

[0114] As shown in Figure 3, CD38 molecules are expressed on the surface of various immune cells, and the expression level of CD38 on the surface of immune cells in ITP patients is significantly higher than that in healthy controls (*: p < 0.05; **: p ≤ 0.01; ***: p ≤ 0.001). As shown in Figure 4, after anti-CD38 monoclonal antibody treatment, the expression level of CD38 on the surface of each immune cell in the patient's peripheral blood was significantly reduced (taking monocytes as an example), and the serum antibody level continued to decrease.

[0115] ITP mouse passive model data:

[0116] As shown in Figure 5, after treating ITP mice with anti-mouse CD38 monoclonal antibody, the platelet count in the tail vein blood routine monitoring was increased, as shown in Figures 6 and 7, after detecting the spleen mononuclear macrophages of mice treated with anti-mouse CD38 monoclonal antibody, it was found that the count of spleen mononuclear macrophages decreased, and the proportion of CD38+ cells decreased significantly (especially macrophages).

[0117] According to the above data, it can be explained that the mechanism of anti-CD38 monoclonal antibody treatment of ITP is to specifically bind CD38 on the surface of mononuclear macrophages, reduce the ADCC function mediated by mononuclear macrophages, and reduce the destruction of platelets. And this mechanism works quickly, and the patient's peripheral platelet count can be significantly improved after receiving 1 drug treatment (1 week of treatment).

[0118] Although the present application has been described in detail with reference to the preferred embodiments, the application is not limited to such but encompasses any modifications or alternatives within the scope of the application as disclosed in the appended claims.

Claims

1. Use of an anti-CD38 antibody in the preparation of a medicament for treating ITP, characterized in that: The anti-CD38 antibody increases the peripheral platelet count by inhibiting the proliferation of mononuclear macrophages.

2. The use according to claim 1, characterized in that The anti-CD38 antibody inhibits the expression of serum antibodies. Preferably, the serum antibodies include IgG, IgM and / or IgA.

3. The use according to any one of claims 1-2, characterized in that The anti-CD38 antibody is an anti-CD38 antibody that can block the activity of mononuclear macrophages.

4. The use according to any one of claims 1 to 3, characterized in that: The ITP includes persistent or chronic ITP.

5. The use according to any one of claims 1 to 4, characterized in that: The ITP patients include patients who have received one or more previous treatments for the first time or in the past. Preferably, the previous treatments include glucocorticoid therapy, immunoglobulin therapy, molecular targeted therapy, thrombocytopoietic drug therapy or immunosuppressant therapy.

6. The use according to any one of claims 1 to 5, characterized in that: The drugs for treating ITP also include methylprednisolone, acetaminophen, antihistamines and / or herpes zoster preventive drugs. The drugs for treating ITP other than the anti-CD38 antibody are administered before, after or simultaneously with the administration of the anti-CD38 antibody.

7. The use according to any one of claims 1 to 6, characterized in that: The administration of the drug includes oral administration, enteral administration, subcutaneous injection, intramuscular injection, intravenous injection, nasal administration, transdermal administration, subconjunctival administration, intraocular administration, orbital administration, retro-ocular administration, retinal administration, choroidal administration, intraperitoneal injection or intrathecal injection.

8. Use of an anti-CD38 antibody in inhibiting the proliferation of monocytes and macrophages.

9. The use according to claim 8, characterized in that The anti-CD38 antibody inhibits the proliferation of mononuclear macrophages, thereby inhibiting the expression of serum antibodies. Preferably, the serum antibodies include IgG, IgM and / or IgA.

10. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises an anti-CD38 antibody and pharmaceutically acceptable excipients. Preferably, the pharmaceutical composition further comprises methylprednisolone, acetaminophen, an antihistamine and / or a herpes zoster preventive drug.

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