CN112741905A - Application of VEGFR inhibitor and anti-PD-1 antibody in preparation of medicine for treating gestational trophoblastic tumor - Google Patents

Abstract

The present disclosure relates to the use of a VEGFR inhibitor in combination with an anti-PD-1 antibody for the preparation of a medicament for the treatment of gestational trophoblastic tumors. Specifically, the anti-PD-1 antibody related to the disclosure is a humanized antibody, and the VEGFR inhibitor is selected from tamifanib, canertib, apatinib and the like or medicinal salts thereof.

Description

Application of VEGFR inhibitor and anti-PD-1 antibody in preparation of medicine for treating gestational trophoblastic tumor

Technical Field

The disclosure relates to an application of a VEGFR inhibitor and an anti-PD-1 antibody in preparation of a medicine for treating gestational trophoblastic tumor.

Background

Gestational Trophoblastic Disease (GTD) is a type of placental trophoblastic disease. The tissue morphology characteristics of the cancer cells include grape embryo, erosive grape embryo, placental site trophoblastic tumor, choriocarcinoma, etc., wherein erosive grape embryo (IM), choriocarcinoma (CC), Placental Site Trophoblastic Tumor (PSTT), and epithelial-like trophoblastic tumor (ETT) are also collectively called gestational trophoblastic tumor (GTN).

Epidemiological survey in China shows that the incidence rate of the grape embryo pregnancy is 0.81 per mill (calculated by thousands of pregnancies); in recent years, the incidence of grape fetuses has decreased, probably due to improvements in economy and diet and decreased fertility. Most of the hydatidiform mole can be cured by clearing the uterus, but some cases can develop into erosive hydatidiform mole. The miscarriage rate of the complete hydatidiform mole is 20 percent, but the miscarriage rate obviously rises when high-risk factors exist, and the high-risk factors of the blood HCG level, the age of a patient and the repeated hydatidiform mole are all the high-risk factors of the miscarriage. The pathological feature of erosive hydatid fetus is that the hydatid fetus tissue erodes the myometrium or other parts, which usually shows symptoms of vaginal bleeding, abdominal pain, abdominal mass, blood HCG overhigh and the like, and the common metastasis parts are lung, brain, nervous system, bladder and the like. The mortality rate of erosive hydatid fetuses is up to 25% before effective chemotherapeutic drugs are discovered, and there are essentially no cases of death since the late 20 th century 50 s when large doses of methotrexate were demonstrated to be effective in treating the tumor and after a series of effective chemotherapeutic drugs were subsequently discovered. For the treatment of erosive hydatidiform mole, reference is now made clinically to the treatment of choriocarcinoma generally.

Choriocarcinoma (choriocarcinoma) is low in incidence, difficult to estimate, about (1-9)/40000 pregnancies, and choriocarcinoma occurring after a grape fetus is difficult to distinguish from erosive grape fetus due to clinical lack of histopathological evidence; placental Site Trophoblastic Tumors (PSTT) and Epithelioid Trophoblastic Tumors (ETT) are much rarer than choriocarcinomas.

The therapeutic principle of GTN is mainly chemotherapy and is assisted by other therapeutic means such as surgery and radiotherapy. For selective low-risk patients with prognosis score of FIGO less than 4, grape fetus in last pregnancy and non-choriocarcinoma in pathological diagnosis, the commonly used first-line single chemotherapeutic drugs comprise MTX, Act-D, 5-Fu and the like. For low-risk patients with prognosis score of 5-6 or choriocarcinoma pathologically diagnosed, the risk of first-line single-drug chemotherapy failure is obviously increased, and combined chemotherapy can be selected according to the scheme of high-risk patients with prognosis score; the high-risk GTN is mainly combined with chemotherapy and is combined with other treatments such as operation and the like for comprehensive treatment. The chemotherapy scheme of the high-risk GTN is firstly extrapolated to an EMA-CO scheme or a combined chemotherapy scheme mainly based on 5-FU, and the complete remission rate and long-term survival rate of the EMA-CO scheme for treating the high-risk metastasis cases for the first time are both more than 90%. The prognosis score is more than or equal to 13, and the second-line treatment scheme of EP-EMA and the like can be directly selected for hepatic brain with poor response to first-line combined chemotherapy or extremely-high-risk trophoblastic tumors with wide metastasis, but the patients can be treated by strong chemotherapy at first, possibly causing hemorrhage, septicemia and even organ failure, and can be treated by low-dose intensity chemotherapy before standard chemotherapy.

Although GTN is highly sensitive to chemotherapy, there is still a 10% drug resistance/recurrence ratio. The diagnostic standard of the drug-resistant GTN generally considers that after continuous chemotherapy for 3 courses, serum HCG is in a platform shape or HCG even rises after 2 courses of treatment, or imaging examination indicates that tumor focus is enlarged or new focus appears, and then the disease is considered as drug-resistant GTN; while there is no accepted diagnostic standard for relapsing GTN for a while, it is generally clinically accepted that elevated HCG levels after one month of discontinuation of treatment suggest relapse. For this group of patients, despite the numerous remedial chemotherapy regimens, there is no clear, highly effective, low toxicity remedial chemotherapy regimen to date. Chemotherapeutic regimens that may be considered include EMA-EP, ICE, VIP, TE/TP, VCR + FUDR + Act-D + VP16, and the like. Arterial infusion chemotherapy can improve the efficacy of drug resistant/relapsing patients. Opportune surgery is also an important treatment for high-risk drug-resistant/recurrent GTN.

WO2018068691A discloses the use of an anti-PD-1 antibody in combination with a VEGFR inhibitor, which may be apatinib mesylate or a pharmaceutically acceptable salt thereof, for the preparation of a medicament for the treatment of cancer, which may be selected from breast cancer, lung cancer, stomach cancer, kidney cancer, liver cancer, melanoma, etc., but does not disclose whether the combination of a VEGFR inhibitor and an anti-PD-1 antibody can be used for the treatment of gestational trophoblastic tumors.

Disclosure of Invention

The present disclosure provides for the use of a VEGFR inhibitor in combination with an anti-PD-1 antibody in the preparation of a medicament for the treatment of gestational trophoblastic tumors (GTN).

The gestational trophoblastic tumor in the uses provided by the present disclosure is selected from erosive mole (IM), choriocarcinoma (CC), Placental Site Trophoblastic Tumor (PSTT), epithelial-like trophoblastic tumor (ETT).

The gestational trophoblastic tumors in the application provided by the disclosure are selected from gestational trophoblastic tumors with a prognosis score of more than or equal to 7 (high risk GTN) of 2000FIGO trophoblastic tumors or drug-resistant/recurrent gestational trophoblastic tumors.

PD-1 antibodies are known, preferably the PD-1 antibody light chain variable region contains the shown in SEQ ID NO. 4, SEQ ID NO. 5 and SEQ ID NO. 6 LCDR1, LCDR2 and LCDR 3.

The heavy chain variable region of the PD-1 antibody comprises HCDR1, HCDR2 and HCDR3 shown in SEQ ID NO 1, SEQ ID NO 2 and SEQ ID NO 3 respectively.

Wherein, the CDR sequences are shown in the following table:

name (R)	Sequence of	Numbering
			HCDR1	SYMMS	SEQID NO：1
HCDR2	TISGGGANTYYPDSVKG	SEQID NO：2
			HCDR3	QLYYFDY	SEQID NO：3
LCDR1	LASQTIGTWLT	SEQID NO：4
			LCDR2	TATSLAD	SEQID NO：5
LCDR3	QQVYSIPWT	SEQID NO：6

Preferably, the anti-PD-1 antibody is a humanized antibody.

In some embodiments, the humanized antibody comprises the light chain variable region set forth in SEQ ID NO. 10 or a variant thereof, preferably having an amino acid change of 0 to 10, more preferably the amino acid change of A43S in the light chain variable region set forth in SEQ ID NO. 10; the humanized antibody comprises the heavy chain variable region of SEQ ID NO. 9 or a variant thereof, preferably having 0-10 amino acid changes, more preferably G44R amino acid changes, in the heavy chain variable region of SEQ ID NO. 9.

The sequences of the heavy and light chain variable regions of the humanized antibody are shown as follows:

heavy chain variable region

EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYMMSWVRQAPGKGLEWVATISGGGANTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARQLYYFDYWGQGTTVTVSS

SEQID NO：9

Light chain variable region

DIQMTQSPSSLSASVGDRVTITCLASQTIGTWLTWYQQKPGKAPKLLIYTATSLADGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQVYSIPWTFGGGTKVEIK

SEQID NO：10

In other embodiments, the humanized antibody comprises the light chain of SEQ ID NO. 8 or a variant thereof, preferably having 0-10 amino acid changes in the light chain variable region, more preferably the amino acid change of A43S; the humanized antibody comprises the heavy chain of SEQ ID NO. 7 or a variant thereof, preferably having 0-10 amino acid changes in the heavy chain variable region, more preferably the amino acid change of G44R.

In another embodiment, the humanized antibody comprises a light chain as set forth in SEQ ID NO. 8 and a heavy chain as set forth in SEQ ID NO. 7.

The sequences of the heavy chain and the light chain of the humanized antibody are shown as follows:

heavy chain

EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYMMSWVRQAPGKGLEWVATISGGGANTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARQLYYFDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK

SEQID NO：7

Light chain

DIQMTQSPSSLSASVGDRVTITCLASQTIGTWLTWYQQKPGKAPKLLIYTATSLADGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQVYSIPWTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

SEQID NO：8

The VEGFR inhibitor described in the present disclosure is selected from the group consisting of PAN-90806, Foretinib, Tafitinib (Tafitinib), Canitinib (Kanitinib), Apatinib (Apatinib), Tanibirumab, Anlotinib (Anlotinib), Delitinib (Lucitinib), Vatalanib, Cediranib (Cediranib), Sevoranib (Chiuranib), Dovirtinib (Dovitinib), Donafenib (Donafenib), Famitininib (Famitininib), Sitratinib, Teratinib (Telatinib), L-21649, TAS-115, Cabovatinib (Cabozantinib), Thioenphenib (Thioenphenib), Fuquanitinib (Fruetinib), Bluenitinib (Brivatib), Brivatinib (Brivatinib), Thionenib (Sufantinib), Sufantinib (Sufanib (Sufantinib), Sufantinib (Sufanib (Sufantinib), Sufantinib (Sufanib (Sufantinib), Sufantini, A pharmaceutically acceptable salt of Rebastinib or a pharmaceutically acceptable salt thereof.

The VEGFR inhibitors provided in the present disclosure in combination with anti-PD-1 antibodies can adjust the dosage regimen to provide the most desirable response, e.g., the maximum therapeutic response and/or the minimum adverse effects.

In some embodiments, the VEGFR inhibitor is apatinib, or a pharmaceutically acceptable salt thereof.

In some embodiments, the dose of apatinib, or a pharmaceutically acceptable salt thereof, is 100-500mg, specifically 100mg, 125mg, 150mg, 175mg, 200mg, 225mg, 250mg, 275mg, 300mg, 325mg, 350mg, 375mg, 400mg, 500mg, preferably 250mg, 375mg, 500mg, once daily, once bidaily, once three days, preferably once daily.

In an alternative embodiment, the apatinib, or pharmaceutically acceptable salt thereof, is administered at a dose of 250mg once a day.

In an alternative embodiment, the apatinib, or pharmaceutically acceptable salt thereof, is administered at a dose of 350mg once a day.

In an alternative embodiment, the apatinib, or pharmaceutically acceptable salt thereof, is administered at a dose of 500mg once a day.

The use of the present disclosure, the dose of the anti-PD-1 antibody administered in a human subject, administered according to the weight of the patient, may be in the following range: 0.1-10.0mg/kg, 0.1-5mg/kg, 1-5mg/kg, 2-5mg/kg, for example, the dose may be 0.1mg/kg, 0.2mg/kg, 0.3mg/kg, 0.4mg/kg, 0.5mg/kg, 0.6mg/kg, 0.7mg/kg, 0.8mg/kg, 0.9mg/kg, 1.0mg/kg, 1.2mg/kg, 1.4mg/kg, 1.6mg/kg, 1.8mg/kg, 2.0mg/kg, 2.2mg/kg, 2.4mg/kg, 2.6mg/kg, 2.8mg/kg, 3.0mg/kg, 3.2mg/kg, 3.4mg/kg, 3.6mg/kg, 3.8mg/kg, 4.0mg/kg, 4.2mg/kg, 4.6mg/kg, 4mg/kg, 4.5 mg/kg, 0mg/kg, 4.8mg/kg, 4mg/kg, 5.2mg/kg, 5.4mg/kg, 5.6mg/kg, 5.8mg/kg, 6.0mg/kg, 6.2mg/kg, 6.4mg/kg, 6.6mg/kg, 6.8mg/kg, 7.0mg/kg, 7.2mg/kg, 7.4mg/kg, 7.6mg/kg, 7.8mg/kg, 8.0mg/kg, 8.2mg/kg, 8.4mg/kg, 8.6mg/kg, 8.8mg/kg, 9.0mg/kg, 9.2mg/kg, 9.4mg/kg, 9.6mg/kg, 9.8mg/kg, 10.0 mg/kg.

In alternative embodiments, wherein the PD-1 antibody is administered in a human subject at a dose within the following range: 1-1000mg, 50-800mg, 50-700mg, 50-600mg, 50-500mg, 50-400mg, 50-300mg, 100-300mg or 200-300 mg. For example, the dose may be 1.0mg, 5.0mg, 10.0mg, 15mg, 20mg, 25mg, 30mg, 35mg, 40mg, 45mg, 50mg, 55mg, 60mg, 65mg, 70mg, 75mg, 80mg, 85mg, 90mg, 95mg, 100mg, 105mg, 110mg, 115mg, 120mg, 125mg, 130mg, 135mg, 140mg, 145mg, 150mg, 155mg, 160mg, 165mg, 170mg, 175mg, 180mg, 185mg, 190mg, 195mg, 200mg, 205mg, 210mg, 215mg, 220mg, 225mg, 230mg, 235mg, 240mg, 245mg, 250mg, 255mg, 260mg, 265mg, 270mg, 275mg, 280mg, 285mg, 290mg, 300mg, 305mg, 310mg, 315mg, 320mg, 325mg, 330mg, 335mg, 350mg, 340mg, 375mg, 370mg, 410mg, 380mg, 390mg, 380mg, 365mg, 380mg, 415mg, 420mg, 425mg, 430mg, 435mg, 440mg, 445mg, 450mg, 455mg, 460mg, 465mg, 470mg, 475mg, 480mg, 485mg, 490mg, 495mg, 500mg, 505mg, 510mg, 515mg, 520mg, 525mg, 530mg, 535mg, 540mg, 545mg, 550mg, 555mg, 560mg, 565mg, 570mg, 575mg, 580mg, 585mg, 590mg, 595mg, 600mg, 605mg, 610mg, 615mg, 620mg, 625mg, 630mg, 635mg, 640mg, 645mg, 650mg, 655mg, 660mg, 665mg, 670mg, 675mg, 680mg, 685mg, 690mg, 695mg, 700mg, 705mg, 710mg, 715mg, 720mg, 725mg, 730mg, 745mg, 735mg, 750mg, 755mg, 760mg, 765mg, 775mg, 770mg, 790mg, 800mg, 830mg, 800mg, 820mg, 800mg, 260mg, 800mg, 260mg, 845mg, 850mg, 855mg, 860mg, 865mg, 870mg, 875mg, 880mg, 885mg, 890mg, 895mg, 900mg, 905mg, 910mg, 915mg, 920mg, 925mg, 930mg, 935mg, 940mg, 945mg, 950mg, 955mg, 960mg, 965mg, 970mg, 975mg, 980mg, 985mg, 990mg, 995mg, 1000mg, preferably 50 to 600mg, most preferably 200 mg.

The anti-PD-1 antibodies of the present disclosure are administered weekly, every 2 weeks, every 3 weeks, every 4 weeks, 1 month, or every 3-6 months, preferably weekly.

In alternative embodiments, the anti-PD-1 antibodies of the present disclosure are administered in a dose of 50-600mg once every 2-3 weeks, preferably 200mg once every 2-3 weeks in a human subject.

In certain embodiments, the anti-PD-1 antibody is administered once every 2 weeks, and in preferred embodiments, the antibody is administered once every 3 weeks.

In certain embodiments, the anti-PD-1 antibody is administered at a dose of 200mg every 2 weeks.

In certain embodiments, the present disclosure provides for the use of apatinib, or a pharmaceutically acceptable salt thereof, in a dose of 250mg, 375mg, 500mg, once daily; the anti-PD-1 antibody was administered at a dose of 200-300mg, at a frequency of once every 2 weeks or once every 3 weeks.

In an alternative embodiment, the present disclosure provides a use of apatinib, or a pharmaceutically acceptable salt thereof, in a dose of 250mg, administered once a day; the anti-PD-1 antibody was administered at a dose of 200mg at a frequency of once every 2 weeks.

In an alternative embodiment, the present disclosure provides a use, wherein the apatinib, or a pharmaceutically acceptable salt thereof, is administered at a dose of 350mg, once a day; the anti-PD-1 antibody was administered at a dose of 200mg at a frequency of once every 2 weeks.

In an alternative embodiment, the present disclosure provides a use, wherein the apatinib, or a pharmaceutically acceptable salt thereof, is administered at a dose of 500mg, once a day; the anti-PD-1 antibody was administered at a dose of 200mg at a frequency of once every 2 weeks.

In certain embodiments, any combination therapy of the present disclosure lasts for at least 2 weeks, or at least 1 month, or at least 3 months, or at least 6 months, or 9 months, or at least 1 year, or at least 18 months, or at least 24 months.

In certain embodiments, the pharmaceutically acceptable salt of apatinib is selected from, but not limited to, the hydrochloride, mesylate, maleate, malate, or besylate salt, and the like, preferably from the mesylate salt.

The present disclosure provides a method of treating a gestational trophoblastic tumor comprising administering to a patient a therapeutically effective amount of apatinib, or a pharmaceutically acceptable salt thereof, and an anti-PD-1 antibody.

The present disclosure provides an anti-PD-1 antibody for use in the treatment of a gestational trophoblastic tumor, wherein the anti-PD-1 antibody is used in combination with a VEGFR inhibitor, wherein the VEGFR inhibitor is preferably apatinib or a pharmaceutically acceptable salt thereof.

The present disclosure provides a VEGFR inhibitor for use in treating a gestational trophoblastic tumor, wherein the VEGFR inhibitor is used in combination with an anti-PD-1 antibody, wherein the VEGFR inhibitor is preferably apatinib or a pharmaceutically acceptable salt thereof.

The route of administration of the combination of the present disclosure is selected from oral, parenteral, transdermal, including but not limited to intravenous, subcutaneous, intramuscular.

The anti-PD-1 antibodies of the present disclosure can be constituted in a composition. In certain embodiments of the present disclosure, the PD-1 antibody is administered by injection, for example, subcutaneously or intravenously or intraperitoneally, and the PD-1 antibody is formulated into an injectable form prior to injection. An injectable form of the anti-PD-1 antibody in a particularly preferred embodiment of the present disclosure is an injection solution or a lyophilized powder injection, which comprises the PD-1 antibody, a buffer, a stabilizer, and optionally further comprising a surfactant. The buffer can be one or more selected from acetate, citrate, succinate and phosphate. The stabilizer may be selected from sugars or amino acids, preferably disaccharides, such as sucrose, lactose, trehalose, maltose. The surfactant is selected from polyoxyethylene hydrogenated castor oil, glycerin fatty acid ester, polyoxyethylene sorbitan fatty acid ester, preferably the polyoxyethylene sorbitan fatty acid ester is polysorbate 20, 40, 60 or 80, most preferably polysorbate 20. The most preferred injectable form of the PD-1 antibody comprises the PD-1 antibody, acetate buffer, trehalose, and polysorbate 20.

Unless otherwise defined, terms in this disclosure have the following meanings:

the "combination" as referred to in the present disclosure is a mode of administration, which means that at least one dose of apatinib or a pharmaceutically acceptable salt thereof and the anti-PD-1 antibody is administered over a certain period of time, wherein both drugs show pharmacological effects. The time period may be within one administration cycle, preferably within 4 weeks, within 3 weeks, within 2 weeks, within 1 week, or within 24 hours. The apatinib, or a pharmaceutically acceptable salt thereof, and the anti-PD-1 antibody may be administered simultaneously or sequentially. Such terms include treatments wherein apatinib, or a pharmaceutically acceptable salt thereof, and the anti-PD-1 antibody are administered by the same route of administration or different routes of administration.

The "effective amount" or "therapeutically effective amount" described in this disclosure includes an amount sufficient to ameliorate or prevent a symptom or condition of a medical condition. An effective amount also means an amount sufficient to allow or facilitate diagnosis. The effective amount for a particular patient or veterinary subject may vary depending on the following factors: such as the condition to be treated, the general health of the patient, the method and dosage of administration, and the severity of side effects. An effective amount may be the maximum dose or dosage regimen that avoids significant side effects or toxic effects.

Objective Response Rate (ORR) refers to the proportion of patients whose tumors have shrunk to a certain extent and are maintained for a certain period of time, including cases of CR and PR. Solid tumor remission evaluation criteria (RECIST 1.1 criteria) were used to assess objective tumor remission. Subjects must be accompanied by measurable tumor lesions at baseline, and the criteria for efficacy assessment are divided into Complete Remission (CR), Partial Remission (PR), Stable (SD), Progression (PD) according to RECIST1.1 criteria.

Disease Control Rate (DCR) refers to the percentage of confirmed cases of complete remission, partial remission, and stable Disease (> 8 weeks) among patients with evaluable efficacy.

Complete Remission (CR): all target lesions disappeared and the short diameter of all pathological lymph nodes (including target and non-target nodes) had to be reduced to < 10 mm.

Partial Remission (PR): the sum of the target lesion diameters is reduced by at least 30% from baseline levels.

Disease Progression (PD): the diameter and relative increase is at least 20% with respect to the minimum of the sum of all measured target lesion diameters throughout the experimental study (baseline values are referenced if the baseline measurement is minimal); in addition to this, it must be satisfied that the absolute value of the sum of the diameters increases by at least 5mm (the appearance of one or more new lesions is also considered as disease progression).

Disease Stability (SD): the target lesion was decreased to a degree that did not reach PR and increased to a degree that did not reach PD levels, between which the minimum of the sum of the diameters was considered for the study.

Detailed Description

The present disclosure is further described below with reference to examples, but these examples do not limit the scope of the present disclosure.

Example 1 effectiveness and safety of PD-1 antibody in combination with Apatinib mesylate for treatment of relapsed/drug resistant gestational trophoblastic tumors

1. Experimental drugs

PD-1 antibody: the commercially available carpolizumab for injection is 200mg, and is a freeze-dried powder injection;

apatinib: commercially available apatinib mesylate tablets.

2. Subjects in group

All of the following inclusion criteria must be met in order to qualify for entry into the test.

(1) (ii) resistant/relapsed GTN patients who failed at least two or more combination chemotherapy regimens;

(2)2000FIGO trophoblastic tumor prognosis score is more than or equal to 7 points (high risk GTN patients) or drug-resistant or recurrent epithelioid trophoblastic tumor or placenta position trophoblastic tumor;

(3)18-70 years old;

(4) patient ECOG score 0-2 points;

(5) abnormal blood HCG levels and/or clinically evaluable lesions with a maximum metastatic focus diameter > 1cm according to iRECIST and RECIST 1.1.

(6) The expected survival time exceeds 4 months;

(7) the main organ function index meets the standard of conventional targeting and immunotherapy

1) Hemoglobin (Hb) is more than or equal to 80g/L,

2) the neutrophil granulocytes (ANC) is more than or equal to 1.5 multiplied by 109/L,

3) platelet count (PLT) is more than or equal to 100X 109/L,

4) serum creatinine (Cr) is less than or equal to 1.5 multiplied by the Upper Limit of Normal (ULN),

5) blood Urea Nitrogen (BUN) is less than or equal to 2.5 multiplied by the Upper Limit of Normal (ULN);

6) total Bilirubin (TB) is less than or equal to ULN;

7) glutamic-oxaloacetic transaminase (AST) and glutamic-pyruvic transaminase (ALT) are less than or equal to 2.5 × ULN;

8) albumin (ALB) is more than or equal to 25 g/L;

9) thyroid Stimulating Hormone (TSH) is less than or equal to the upper limit of normal value (ULN); if the T3 and T4 levels should be considered abnormally, the T3 and T4 levels are normal and can be selected.

(8) Female subjects of child bearing age must exclude pregnancy and would like to use a medically approved high-performance contraceptive (e.g., an intrauterine device, contraceptive, or condom) during the study and within 3 months after the last administration of study medication

(9) Informed consent was signed and subjects were informed of the purpose of the study and the procedures required for the study, and were voluntarily enrolled in the study.

3. Method of administration

PD-1 antibody, intravenous drip, fixed dose 200 mg/time. Intravenous drip for 30min (not less than 20min and not more than 60min) every 2 weeks;

apatinib tablet, 250mg, is orally administered 1 time per day. Is taken after half an hour.

The combination regimen was continued until the following events occurred:

end of combined maintenance treatment half a year after the subject reached Complete Remission (CR);

failure of the subject to remission or disease progression;

intolerance of toxicity;

or other reasons for protocol specification;

4. results of the experiment

The group is 11, 6 cases are evaluated for the first time, 5 cases are PR, and 1 case is SD.

The late study included a total of 20 drug resistant/relapsed GTN patients. In terms of therapeutic efficacy, the patients had an ORR rate of 55% (11/20), with a CR rate of 50% (10/20) and a Partial Remission (PR) rate of 5% (1/20). In terms of safety, the most common adverse effect was neutropenia (40%), and there were 12 patients who developed 3/4-grade treatment-related adverse effects during the treatment period, mainly including hypertension, rash, leukopenia, elevated transaminase, neutropenia, and hand-foot syndrome, and no treatment-related death occurred during the treatment period.

Sequence listing

<110> Hengrui pharmaceutical Co., Ltd of Jiangsu

SUZHOU SUNCADIA BIOPHARMACEUTICALS Co.,Ltd.

SHANGHAI HENGRUI PHARMACEUTICAL Co.,Ltd.

Application of VEGFR inhibitor and anti-PD-1 antibody in preparation of medicine for treating gestational trophoblastic tumor

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<210> 1

<211> 5

<212> PRT

<213> mouse source (Mus musculus)

<400> 1

Ser Tyr Met Met Ser

1 5

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<211> 17

<212> PRT

<213> mouse source (Mus musculus)

<400> 2

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

1 5 10 15

Gly

<210> 3

<211> 7

<212> PRT

<213> mouse source (Mus musculus)

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Gln Leu Tyr Tyr Phe Asp Tyr

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<213> mouse source (Mus musculus)

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Leu Ala Ser Gln Thr Ile Gly Thr Trp Leu Thr

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<213> mouse source (Mus musculus)

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Thr Ala Thr Ser Leu Ala Asp

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<213> mouse source (Mus musculus)

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Gln Gln Val Tyr Ser Ile Pro Trp Thr

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<210> 7

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<220>

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<222> (1)..(443)

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

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

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

35 40 45

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

50 55 60

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

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

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Ala Arg Gln Leu Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Thr Val

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

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Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu

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Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

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

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Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

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

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Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro

210 215 220

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

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

245 250 255

Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn

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

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

290 295 300

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

305 310 315 320

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

325 330 335

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

340 345 350

Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe

355 360 365

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

370 375 380

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

385 390 395 400

Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly

405 410 415

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

420 425 430

Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 8

<211> 214

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<221> PEPTIDE

<222> (1)..(214)

<223> light chain sequence

<400> 8

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 Leu Ala Ser Gln Thr Ile Gly Thr Trp

20 25 30

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

35 40 45

Tyr Thr Ala Thr Ser Leu Ala Asp 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 Val Tyr Ser Ile Pro Trp

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

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

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

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

180 185 190

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

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 9

<211> 116

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<221> PEPTIDE

<222> (1)..(116)

<223> heavy chain variable region

<400> 9

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 Tyr

20 25 30

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

35 40 45

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

50 55 60

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

100 105 110

Thr Val Ser Ser

115

<210> 10

<211> 107

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<221> PEPTIDE

<222> (1)..(10)

<223> light chain variable region

<400> 10

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 Leu Ala Ser Gln Thr Ile Gly Thr Trp

20 25 30

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

35 40 45

Tyr Thr Ala Thr Ser Leu Ala Asp 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 Val Tyr Ser Ile Pro Trp

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

Claims (12)

1. Use of a VEGFR inhibitor in combination with an anti-PD-1 antibody in the preparation of a medicament for the treatment of gestational trophoblastic tumors.

2. The use according to claim 1, wherein said gestational trophoblastic tumor is selected from the group consisting of erosive hydatidiform mole, choriocarcinoma, placental site trophoblastic tumor, epithelioid trophoblastic tumor.

3. The use of claim 1, wherein the gestational trophoblastic tumor is selected from a 2000FIGO trophoblastic tumor prognosis score of 7 points or more (high risk GTN) or a drug-resistant/relapsed gestational trophoblastic tumor.

4. The use according to any one of claims 1 to 3, wherein the light chain variable region of the anti-PD-1 antibody comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO 4, SEQ ID NO 5 and SEQ ID NO 6, respectively, and the heavy chain variable region of the PD-1 antibody comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO 1, SEQ ID NO 2 and SEQ ID NO 3, respectively.

5. The use of claim 4, wherein the anti-PD-1 antibody is a humanized antibody.

6. The use of claim 5, wherein the humanized antibody comprises a light chain variable region as set forth in SEQ ID No. 10 or a variant thereof, preferably having an amino acid change of 0 to 10, more preferably an amino acid change of A43S in the light chain variable region as set forth in SEQ ID No. 10; the humanized antibody comprises the heavy chain variable region of SEQ ID NO. 9 or a variant thereof, preferably having 0-10 amino acid changes, more preferably G44R amino acid changes, in the heavy chain variable region of SEQ ID NO. 9.

7. The use according to claim 5, wherein the humanized antibody comprises the light chain of SEQ ID NO 8 or a variant thereof, preferably having 0-10 amino acid changes in the light chain variable region, more preferably the amino acid change of A43S; the humanized antibody comprises the heavy chain of SEQ ID NO. 7 or a variant thereof, preferably having 0-10 amino acid changes in the heavy chain variable region, more preferably the amino acid change of G44R.

8. The use of claim 7, wherein the humanized antibody comprises a light chain as set forth in SEQ ID NO 8 and a heavy chain as set forth in SEQ ID NO 7.

9. The use according to any one of claims 1 to 8, wherein the VEGFR inhibitor is selected from the group consisting of PAN-90806, Foretinib, Tafitinib (Tafetinib), canertinib (Kanitiib), Apatinib (Apatinib), Tanibirumab, Anlotinib (Anlotinib), Delitinib (Lucitinib), Vatalanib, Cediranib (Cediraniib), Seaoraniib (Chiauraraniib), Durvertinib (Dovitib), Dunnafenib (Donafenib), Famitatinib (Famitatinib), Sivatinib (Terlatinib), L-21649, TAS-115, Cabozantinib (Cabozantinib), Thielafenib (Thiophenib), Furofenib (Fukulitinib), Britisib (Tenuitinib), Brininib (Brininib), Brinilotinib (Brininib), Veninib (Sunifibrib), Vernib (Sunifibrib (Piraninib), Vernib (Sunifibrib (Edoniib), Vernib (Piraninib), Vernib (Sunifibrib), Vernib (Sunifibrib), Vernib (Sunifibrib (, Altiratinib, Ningetinib (Nigertinib), Sunitinib (Sunitinib), AL-8326, Rebastinib or pharmaceutically acceptable salts thereof.

10. Use according to claim 9, wherein the pharmaceutically acceptable salt of apatinib is selected from the group consisting of the hydrochloride, mesylate, maleate, malate or besylate salt, preferably from the mesylate salt.

11. The use according to claim 9 or 10, wherein the dose of apatinib or a pharmaceutically acceptable salt thereof is 100-500mg, preferably 250mg, 375mg, 500 mg; the administration frequency is once a day, once every two days, once every three days, preferably once a day.

12. The use according to any one of claims 1 to 11, the anti-PD-1 antibody dose is selected from 0.1-10.0mg/kg or 1-1000mg, preferably 200 mg; the frequency of administration is weekly, every 2 weeks, every 3 weeks or every 4 weeks, preferably every 2 weeks or every 3 weeks.