Application of neutrophil number level in prediction of tumor treatment effect

Abstract

The invention discloses application of neutrophile granulocyte number level in prediction of tumor treatment effect. Through verification of the embodiment of the invention, the number level of neutrophils in blood of a tumor patient before treatment is closely associated with the curative effect of tumor treatment, especially OH2 treatment, and the applicable tumor adaptation diseases are common; comprising at least one of melanoma, esophageal cancer, right breast invasive ductal cancer, intestinal cancer, head and neck tumor, gastric cancer, ovarian cancer, biliary system tumor, left foot back squamous cell carcinoma, sarcoma and osteosarcoma, lung cancer, thymosquamous cell carcinoma and brain glioma. In the application disclosed by the invention, the threshold value of the number of neutrophils for predicting the tumor treatment effect is 3.5 * 10 < 9 > / L; the treatment is predicted to be effective when the number of sexual granulocytes in a tumor patient is below 3.5 * 109 / L before the tumor patient is treated, and the treatment is predicted to be ineffective or invalid when the number of sexual granulocytes in the tumor patient is higher than 3.5 * 109 / L. The application method is simple, low in cost and good in stability, and can become a potential prediction means for clinicians.

Description

Technical Field

[0001] The present application relates to the technical field of tumor efficacy prediction, and specifically to an application based on neutrophil count levels in predicting tumor treatment efficacy. Background Art

[0002] Cancer is an important disease that affects and endangers people's physical health and life safety, and has long been ranked in the top two in disease mortality. Cancer also belongs to a large group of diseases that can arise from almost any organ or tissue in the body. Abnormal cells grow uncontrollably, divide and grow without limit, exceed their usual boundaries, invade adjacent parts of the body and / or spread to other organs. Tumors and malignant tumors are other common names for cancer. There are many types of tumors, which can be divided into brain tumors, liver tumors, digestive tract tumors, melanoma, etc. according to the site of onset. There are many ways to treat tumors, the most common ones are surgical treatment, radiotherapy, chemotherapy, molecular targeted therapy and immunotherapy.

[0003] Oncolytic virus therapy is a new anti-tumor method that uses viruses to attack cells to destroy tumor tissue. Oncolytic virus is a virus that can infect and destroy tumor cells. Oncolytic virus therapy has broad application prospects and development potential, and more and more evidence supports it as an effective anti-tumor treatment. Recombinant human GM-CSF oncolytic herpes simplex virus type II (OH2) injection (Vero cells) is a new type of tumor vaccine and tumor gene therapy drug of Class I. This type of oncolytic virus drug not only replicates and lyses tumor cells in large quantities in the tumor, but also effectively activates the immune system including T cells, forming a dual anti-tumor effect with a broad-spectrum, potent and low-toxic anti-tumor effect. Its treatment is mainly aimed at solid tumors, including lung cancer, head and neck tumors, lymphoma, liver cancer, melanoma, etc.

[0004] With the gradual deepening of clinical research, OH2 injection is increasingly used in the treatment of tumors. Tumor immunotherapy is affected by many factors, which also leads to differences in the therapeutic efficacy of OH2 injection in different tumor patients. Therefore, OH2 treatment requires efficient and convenient biomarkers to timely and accurately identify the responding population and avoid or reduce unnecessary economic burdens on the non-responding population.

[0005] In summary, there is still a lack of accurate and efficient markers in clinical practice to predict whether OH2 is effective in treating tumors. Summary of the invention

[0006] In view of this, the purpose of the present application is to provide an application based on the neutrophil count level in predicting the efficacy of tumor treatment. In this application, by testing the neutrophil count level of tumor patients, it is not only helpful to more accurately screen the group of tumor patients suitable for OH2 treatment, but also to provide a scientific basis for optimizing treatment plans and improving treatment effects.

[0007] In order to achieve or at least partially achieve the above objectives, this application provides at least the following technical solutions:

[0008] An application based on neutrophil count level in predicting the efficacy of tumor treatment, wherein the neutrophil count level is the neutrophil count level in the blood of tumor patients before tumor treatment.

[0009] Furthermore, in the application, the tumor treatment method is to receive oncolytic virus drug treatment.

[0010] Preferably, in the application, the tumor treatment method is to receive treatment with OH2 virus or OH2 virus preparation.

[0011] Furthermore, in the application, the threshold value of neutrophil count for predicting the efficacy of tumor treatment is 3.5×10 9 / L; before receiving treatment, the neutrophil count of cancer patients is 3.5×10 9 / L or below, predicting treatment effectiveness, when the neutrophil count is higher than 3.5×10 9 / L, it is predicted that the treatment is ineffective or ineffective.

[0012] Furthermore, in the application, the applicable tumor types include at least one of melanoma, esophageal cancer, right breast invasive ductal carcinoma, intestinal cancer, head and neck tumors, gastric cancer, ovarian cancer, biliary system tumors, left dorsal squamous cell carcinoma, sarcoma and osteosarcoma, lung cancer, thymic squamous cell carcinoma, and brain glioma.

[0013] Compared with the prior art, the technical solution provided by this application has at least the following beneficial effects:

[0014] 1. Through verification by the examples of the present application, the neutrophil count level of tumor patients before treatment is closely related to the efficacy of tumor treatment. Therefore, the neutrophil count level can be used as an efficacy prediction factor to more accurately screen the patient group suitable for OH2 treatment, providing a scientific basis for optimizing the treatment plan and improving the treatment effect.

[0015] 2. The technical solution provided in this application has a simple detection method, low cost, and good stability, and can become a potential predictive tool for clinicians. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1Representative box plots of immune cell counts of subjects in the clinical benefit group and clinical non-benefit group in the discovery set provided in the examples of the present application. DETAILED DESCRIPTION

[0017] In order to make the purpose, technical solution and advantages of the present application more clearly understood, the present application is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present application and are not used to limit the present application.

[0018] In the present application text, the term "OH2" refers to "OH2 virus", the full name of which is "recombinant oncolytic herpes simplex virus type II", which uses molecular cloning, DNA (not evaluated) homologous recombination and other technologies to delete the neurotoxic gene ICP34.5 gene and the immunosuppressive gene ICP47 gene in the viral genome, and insert a human hGMCSF expression cassette in the ICP34.5 region. The gene nucleotide sequence of the expression cassette is shown in SEQ ID NO: 1. The "OH2 virus" strain is named H2d3d4-hGF, and the recommended classification name is: herpes simplex virus type II (Latin name: Herpes Simplex Virus Type 2). The depository is the General Microbiology Center of the China Microbiological Culture Collection Administration, located at the Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC No. 3600, and the deposit date is February 3, 2010.

[0019] In the text of this application, the term "injection preparation" refers to a sterile solution (including emulsions and suspensions) made of medicine for injection into the body, as well as a sterile powder or concentrated solution for preparation into a solution or suspension before clinical use. Injections act quickly and reliably, are not affected by pH, enzymes, food, etc., have no first-pass effect, can exert systemic or localized effects, and are suitable for patients who are not suitable for oral medication and cannot take oral medication. It should be understood that in the text of this application, the terms "injection" and "injection preparation" are equivalent in definition, and the "OH2 injection" consists of "OH2 virus" particles and a buffer that preserves the "OH2 virus".

[0020] In the text of this application, the term "OH2 treatment" should be understood as treatment with "OH2 virus" or "OH2 virus preparation", such as treatment with "OH2 injection".

[0021] In the text of this application, the term "iRECIST" is the standard for evaluating the efficacy of immunotherapy for solid tumors (the evaluation standard for glioma is iRANO), the term "iSD" indicates stable disease due to immune efficacy, the term "iPR" indicates partial remission due to immune efficacy, the term "iCR" indicates complete remission due to immune efficacy, and the term "iUPD" indicates disease progression to be confirmed.

[0022] In the text of this application, the term "iORR" is an indicator related to the evaluation of the therapeutic efficacy of tumors (solid tumors only), and "objective response rate (iORR)" refers to the sum of the proportions of partial remission (iPR) and complete remission (iCR) of immune efficacy; the higher the iORR, the more patients achieve tumor shrinkage using this immunotherapy method.

[0023] In the text of this application, the term "preferred" is only used to describe an implementation method or example with better effects, and it should be understood that it does not constitute a limitation on the scope of protection of the present invention.

[0024] The present application embodiment provides a clinical application test of neutrophil count level in predicting the efficacy of OH2 in treating tumors. Two clinical sequences of immunotherapy were established during the entire clinical trial process, namely, a discovery set (N1=44) and a validation set (N2=99). The clinical trial process can be divided into two steps: the first step is to track the effect of tumor immunotherapy by detecting the neutrophil count level of 44 melanoma tumor subjects in the discovery set, and screen out markers and detection critical values ​​for predicting the efficacy of OH2 in treating tumors; the second step is to verify the screened markers and detection critical values ​​through the validation set.

[0025] In the present application examples, the inclusion criteria and exclusion criteria for the clinical study cohort subjects are as follows:

[0026] 1. Selection criteria

[0027] (1) Aged 18 to 75 years (including the boundary value), male or female; (2) Patients with malignant tumors diagnosed by pathology or cytology, such as digestive system tumors, head and neck tumors, soft tissue sarcoma, melanoma, etc.; (3) Patients with advanced stage (unresectable or recurrent / metastatic) after failure of standard treatment (disease progression or intolerance) or lack of effective treatment methods; (4) General condition score ECOG 0-1 points; (5) Expected survival period is more than 3 months; (6) Previous anti-tumor treatment (including endocrine, chemo / radiotherapy, targeted therapy) has been completed for more than 4 weeks (for chemotherapy with nitrosoureas and mitomycin, the drug should be discontinued for more than 6 weeks), and the adverse reactions of the previous treatment have recovered to grade 1; (7) Patients who have undergone major surgery must wait for 4 weeks after surgery; (8) At least one measurable or evaluable lesion; (9) There is a lesion suitable for intratumoral injection (the long diameter of the lesion is at least greater than or equal to 0.5 cm); (10) Patients with asymptomatic central nervous system metastases or asymptomatic brain metastases after treatment must have no disease progression by computed tomography (CT) or magnetic resonance imaging (MRI), be stable for at least 3 months, and do not require steroid treatment for at least 4 weeks; (11) Laboratory examination: WBC ≥ 3.0 × 10 9 / L, ANC≥1.5×10 9 / L,PLT≥100×10 9 / L, Hb ≥ 90g / L; blood BUN and serum creatinine within 1.5 times the upper limit of normal; TBIL ≤ 1.5 times the upper limit of normal; ALT and AST ≤ 2.5 times the upper limit of normal; patients with liver metastasis do not exceed 5 times the upper limit of normal; coagulation function is normal (PT, APTT within 1.5 times the upper limit of normal); (12) female subjects and their spouses receive effective contraceptive methods during treatment and within 3 months after treatment; (13) subjects with herpes need to wait for 3 months after the end of herpes; (14) voluntarily sign the informed consent form, and the expected compliance is good.

[0028] In addition to the above conditions, the inclusion criteria for subjects with glioma should also meet the following inclusion criteria:

[0029] (1) Phase I trials select patients who have undergone surgery for pathologically confirmed recurrent central nervous system tumors (including but not limited to anaplastic astrocytoma, anaplastic oligoastrocytoma, glioblastoma, gliosarcoma, anaplastic ependymoma, medulloblastoma, malignant meningioma, melanoma, etc.); Phase IIa trials select patients who have undergone surgery for pathologically confirmed recurrent glioblastoma; (2) KPS score ≥ 60; (3) Partial or complete resection is achieved through surgery, and an Ommaya capsule has been placed in the surgical area, or no substantial resection is performed but an Ommaya capsule can be and has been placed in the surgical area, and the conditions for drug administration are met; (4) According to the investigator's assessment, the expected injection site of the trial drug is in the supratentorial area.

[0030] 2. Exclusion criteria:

[0031] (1) Subjects with serious medical diseases, including serious heart disease, cerebrovascular disease, uncontrolled diabetes, uncontrolled hypertension, severe infection, and active gastrointestinal ulcers; (2) Subjects with clinical symptoms of brain metastases; (3) Subjects with active infection or unexplained fever >38.5°C during the screening period and before the first dose; (4) Subjects with congenital or acquired immune deficiency (such as HIV infection) or active hepatitis (Hepatitis B reference: HbsAg, Anti-HBs, HBeAg, Anti-HBc, Anti-HBe, HBV DNA (not evaluated) ≥10 / mL, hepatocyte transaminase, etc.; Hepatitis C reference: HCV antibody and HCV (5) The lesion cannot meet the requirement of 1 mL injection volume in the tumor; (6) Pregnant or lactating women; (7) Patients who have used or are using other experimental drugs or antiviral treatments within 4 weeks before treatment; (8) Participated in other clinical studies within the past 4 weeks; (9) Patients who are allergic to herpes viruses and drug ingredients; (10) Subjects with a history of psychotropic drug abuse, alcoholism or drug abuse; (11) Patients with other malignant tumors within 5 years before enrollment, excluding effectively resected cervical carcinoma in situ, low-risk gastrointestinal stromal exclusion standard tumors, breast cancer, basal cell carcinoma of the skin, squamous cell carcinoma of the skin, and papillary thyroid carcinoma; (12) Patients with active autoimmune diseases or a history of autoimmune diseases that may relapse, but patients with the following diseases are not excluded and can be further screened: a. Type I diabetes, b. Hypothyroidism (if it can be controlled with hormone replacement therapy alone), c. Controlled celiac disease, d. Skin diseases that do not require systemic treatment (such as vitiligo, psoriasis, alopecia), e. . Any other disease that will not recur without external triggers; (13) Any condition that requires systemic treatment with corticosteroids (prednisone >10 mg / day or equivalent dose of similar drugs) or other immunosuppressants within 14 days before study drug administration, but patients who are currently or have previously used any of the following steroid regimens can be included: a. Epinephrine replacement steroids (prednisone ≤10 mg / day or equivalent dose of similar drugs), b. Topical, ophthalmic, intra-articular, intranasal or inhaled corticosteroids with minimal systemic absorption, c. Prophylactic short-term (≤7 days) use of corticosteroids (e.g., allergy to contrast agents) or for the treatment of non-autoimmune diseases (e.g., delayed hypersensitivity reactions caused by contact allergens); (14) For the OH2 injection and HX008 combination group, subjects who have previously been treated with PD-1 / PD-L1 monoclonal antibodies and have permanently discontinued the drug due to serious adverse reactions; (15) The investigator believes that the patient is unsuitable for participation in this trial for any reason.

[0032] In the present application example, 22 melanoma subjects in the discovery set were divided into a clinical benefit group (defined as patients who received OH2 treatment for ≥12 months) and a clinical non-benefit group (received OH2 treatment for <12 months), and blood samples of these subjects were collected before and after treatment. The Olink (Uppsala, Sweden) proteomics platform was used for retrospective analysis. The results showed that in the clinical non-benefit cohort, several cytokines such as IFN-γ and IL-18 were significantly higher than those in the clinical benefit group at baseline (before treatment) and after treatment. GO enrichment analysis of upregulated proteins showed the involvement of pathways such as neutrophil chemotaxis and cytokine activity. Therefore, in order to test whether the level of neutrophil count is related to the efficacy of OH2 in treating tumors, the present application example analyzed the correlation between the peripheral blood neutrophil count (before treatment) and the efficacy of OH2 treatment in 44 subjects in the discovery set. The results of neutrophil count and efficacy are shown in Table 1 below.

[0033] Table 1

[0034]

[0035]

[0036] Figure 1 Representative box plots of immune cell counts of subjects in the clinical benefit group and the clinical non-benefit group are shown in the figure. As shown in the figure, the baseline level of neutrophil count in the clinical non-benefit group (mean value of 3.93×10 9 / L), which was significantly higher than the baseline level of neutrophil count in the clinical benefit group (mean value was 3.16×10 9 / L), while there was no significant difference in the baseline levels of other lymphocytes and Treg cells.

[0037] The present application example further analyzes the average baseline neutrophil count level in peripheral blood of 44 patients grouped according to different treatment time periods, as shown in Table 2.

[0038] Table 2

[0039]

[0040]

[0041] NR*: Not Reached, indicating that the median survival time has not been reached.

[0042] From the statistical results in Table 2, it can be seen that even if the 44 subjects were grouped according to different treatment (dosing) times, the average baseline neutrophil count in the long-term dosing group was significantly lower than that in the short-term dosing group, and the median overall survival (mOS) of the subject group with a lower baseline neutrophil count level was significantly higher than that of the subject group with a higher baseline neutrophil count level. As can be seen from Table 2, the candidate critical value (threshold) of neutrophil count is set at 3.5×10 9 / L can predict the efficacy of OH2 in the treatment of melanoma. The baseline neutrophil count level is 3.5×10 9 Subjects with blood pressure below 1 / L showed better efficacy after OH2 treatment.

[0043] In order to further verify the above conclusion, the present application example statistically analyzed the neutrophil count level, mOS and iORR of subjects who received OH2 treatment for more than 1 month from the results in Table 1, as shown in Table 3.

[0044] Table 3

[0045]

[0046] As can be seen from Table 3, the 24 neutrophil count levels were ≤3.5×10 9 For subjects with neutrophil counts >3.5×10 / L, the median overall survival (mOS) was not reached but was at least 30.3 months, and the iORR was 29.2%. 9 For subjects with 100 μg / L, the median overall survival (mOS) was not reached, but was at least 22.6 months, and the iORR was 16.7%. Table 4 shows the average baseline neutrophil count level of subjects grouped by the best efficacy.

[0047] Table 4

[0048] Best response (iRECIST) <![CDATA[Average baseline neutrophil count (10 9 / L)]]> iPR (n=10) 3.1 iSD (n=15) 3.6 iUPD (n=17) 4.0 Not evaluated (n=2) 5.4

[0049] As can be seen from Table 4, when the neutrophil count in the blood of the subjects was lower than 3.5×10 9 / L, the subjects could benefit more from OH2 treatment.

[0050] Based on the above-screened markers and critical value levels for predicting the efficacy of tumor treatment, the examples of the present application collected the results of the neutrophil count levels and therapeutic efficacy of subjects with different types of solid tumors (validation set) before treatment (baseline), and conducted verification with an expanded sample size. The baseline neutrophil count levels and therapeutic efficacy results of the validation set subjects are shown in Table 5 below.

[0051] Table 5

[0052]

[0053]

[0054] The results in Table 5 were statistically analyzed to obtain the neutrophil count levels, mOS, and iORR of the 99 validation set subjects, as shown in Table 6.

[0055] Table 6

[0056]

[0057] From the results in Table 6, we can see that the neutrophil count level of 42 patients was ≤3.5×10 9 For subjects with neutrophil counts >3.5×10 / L, the median overall survival (mOS) was 10.4 months and the iORR was 10.5%. 9 For subjects with neutrophil counts of 3.5×10 / L, the median overall survival (mOS) was 5.4 months and the iORR was only 2.2%, indicating that the neutrophil count level was 3.5×10 9 Subjects with other solid tumors below 400 μg / L showed better efficacy after OH2 treatment.

[0058] The neutrophil count levels and efficacy results of all subjects (n=143) in Tables 1 and 6 were statistically analyzed, and mOS and ORR were calculated, as shown in Table 7.

[0059] Table 7

[0060]

[0061] From the results in Table 7, it can be seen that the neutrophil count level of 66 patients was ≤3.5×10 9 For subjects with neutrophil counts >3.5×10 / L, the median overall survival (mOS) was 27.4 months and the iORR was 17.7%. 9 For subjects with neutrophil counts of 3.5×10 / L, the median overall survival (mOS) was 7.9 months and the iORR was only 6.3%, which further indicated that the neutrophil count value was 3.5×10 9 Subjects with tumors below 400 μg / L showed better efficacy after OH2 treatment.

[0062] The above results indicate that there is a close correlation between the level of neutrophils in the blood of cancer patients before treatment and the efficacy of OH2. The reason is that, on the one hand, in the clinical non-benefiting patient group, the increase in the level of neutrophils is accompanied by an increase in the number of polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs). These polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs) have a strong immunosuppressive function and can inhibit T cell function, thereby weakening the anti-tumor efficacy of OH2; on the other hand, high levels of neutrophils can inhibit the replication of OH2 by secreting excessive INF-γ, thereby limiting the effect of OH2 in treating tumors.

[0063] In summary, neutrophils are a key indicator of systemic inflammatory response, and their levels are closely related to tumor development and prognosis. They can be used as a marker to predict the efficacy of OH2 treatment in tumor patients. The critical value (threshold) for prediction is 3.5×10 9 / L, that is, the level of neutrophils in the blood is 3.5×10 9 The efficacy of OH2 treatment in subjects with tumors below 3.5×10 / L was significantly better than that in subjects with neutrophil counts above 3.5×10 9 / L tumor subjects receiving OH2 treatment efficacy.

[0064] The present application is described in detail above. Specific examples are used in this article to illustrate the principles and implementation methods of the present application. The description of the above embodiments is only used to help understand the present application and its core ideas. It should be pointed out that for ordinary technicians in this technical field, without departing from the principles of the present application, several improvements and modifications can be made to the present application, and these improvements and modifications also fall within the scope of protection of the claims of the present application.

Claims

1. An application of neutrophil count level in predicting the therapeutic efficacy of tumors, characterized in that: The neutrophil count level is the neutrophil count level in the blood of tumor patients before tumor treatment.

2. The use according to claim 1, characterized in that: In the application, the tumor is treated by oncolytic virus drug therapy.

3. The use according to claim 1, characterized in that: In the application, the tumor treatment method includes receiving treatment with OH2 virus or OH2 virus preparation.

4. The use according to claim 1, characterized in that: In the application, the threshold of neutrophil count for predicting the efficacy of tumor treatment is 3.5×10 9 / L; before receiving treatment, the neutrophil count of cancer patients is 3.5×10 9 / L or below, predicting treatment effectiveness, when the neutrophil count is higher than 3.5×10 9 / L, it is predicted that the treatment is ineffective or ineffective.

5. The use according to claim 1, characterized in that: In the application, the applicable tumor types include at least one of melanoma, esophageal cancer, right breast invasive ductal carcinoma, intestinal cancer, head and neck tumors, gastric cancer, ovarian cancer, biliary system tumors, left dorsal squamous cell carcinoma, sarcoma and osteosarcoma, lung cancer, thymic squamous cell carcinoma, and brain glioma.

6. An application of neutrophil count level in predicting the efficacy of tumor treatment, characterized in that: In the application, the sample used to detect the level of neutrophil count is peripheral blood of tumor patients.

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