Use of neutrophil count in predicting efficacy of oncolytic virus therapy for tumour

Abstract

Provided is a method of oncolytic virus therapy for a tumour based on using neutrophil count as a predictor of the efficacy of oncolytic virus therapy for the tumour, comprising: determining the neutrophil count level in the blood of a tumour patient prior to tumour treatment; and, when the neutrophil count level in the blood of the tumour patient is below a cut-off value, administering an oncolytic virus to the tumour patient for treatment, wherein the cut-off value is any value from 2×109 to 5.5×109 / L.

Description

Application of neutrophil count in predicting the efficacy of oncolytic virus therapy for tumors

[0001] Cross-reference to related applications

[0002] This application is based on and claims priority to Chinese patent applications No. 202411172772.3, filed on August 26, 2024; No. 202510166466.7, filed on February 14, 2025; and No. 202510633072.8, filed on May 16, 2025, the entire contents of which are incorporated herein by reference. Technical Field

[0003] This disclosure relates to the field of tumor efficacy prediction technology, specifically to the application of neutrophil count in predicting the efficacy of oncolytic virus therapy for tumors. Background Technology

[0004] Cancer is a major disease affecting and endangering people's health and lives, consistently ranking among the top two causes of death. Cancer is also a broad group of diseases that can arise in almost any organ or tissue of the body. Abnormal cells grow uncontrollably, dividing and expanding without restraint, exceeding their normal boundaries, invading adjacent parts of the body and / or spreading to other organs. Tumors and malignant tumors are other common names for cancer. There are many types of tumors, which can be classified according to their location, such as brain tumors, liver tumors, digestive tract tumors, and melanomas. There are many ways to treat cancer, the most common being surgery, radiation therapy, chemotherapy, molecular targeted therapy, and immunotherapy.

[0005] Oncolytic virus (OV) therapy is a novel anti-tumor approach that utilizes viruses to attack cells and destroy tumor tissue. Oncolytic viruses are viruses capable of infecting and destroying tumor cells. They selectively target and lyse tumor cells while preserving normal cells. The anti-tumor effect of OV depends on the induction of local and systemic immune responses. OV preferentially replicates in malignant cells, ultimately inducing tumor cell death. Furthermore, virus-mediated lysis releases tumor-associated antigens, which very effectively stimulate the immune system, thereby promoting the induction of anti-tumor immunity. While some viruses are naturally oncolytic, most OVs are genetically engineered to increase tumor selectivity and improve efficacy. More than 40 OVs from at least 10 different viral families have entered clinical development. In a phase Ib trial for the treatment of advanced melanoma, OV combined with immune checkpoint inhibitors (ICIs) significantly increased CD8+. +T-cell infiltration and response rates. However, an international randomized, double-blind, multicenter phase III trial from J Clin Oncol showed that, compared with placebo, the addition of T-VEC (a recombinant herpes simplex virus-1 approved by the FDA and other regulatory agencies) to the programmed death receptor 1 (PD-1) inhibitor did not significantly improve patient survival outcomes.

[0006] OH2 is a recombinant herpes simplex virus type 2 (HSV-2) that selectively replicates in tumor cells. The OH2 virus removes the ICP34.5 and ICP47 genes from wild-type HSV-2 to attenuate virulence and enhance oncolytic activity. In addition to these deletions, a human GM-CSF gene expression cassette is inserted into the OH2 virus, mediating the expression of the GM-CSF protein in the tumor microenvironment, thereby enhancing the host's immune response. The OH2 virus differs from T-VEC; its backbone is derived from HSV-2. Preclinical studies have shown that in several tumor-bearing animal models, including breast cancer, colon cancer, and metastatic ovarian cancer, HSV-2 (similar to OH2) has better tumor-suppressive effects than HSV-1 (similar to T-VEC). Therefore, OH2 has made progress in clinical trials and is currently undergoing late-stage clinical trials. Phase Ia / Ib clinical trials have been conducted in China for patients with unresectable stage III-IV melanoma, most of whom have acral melanoma, to evaluate the safety and preliminary efficacy of OH2 oncolytic virus therapy. OH2 oncolytic virus therapy has demonstrated good safety with no dose-limiting toxicities. It has also shown durable antitumor efficacy in melanoma patients, particularly those whose disease has progressed after PD-1 therapy.

[0007] As clinical research progresses, the application of OH2 oncolytic viruses in cancer treatment is becoming increasingly widespread, primarily targeting patients with solid tumors, including melanoma, colorectal cancer, glioma, esophageal cancer, head and neck tumors, gastric cancer, biliary system tumors, and sarcomas. Cancer immunotherapy is influenced by multiple factors, leading to variations in the efficacy of OH2 oncolytic viruses among different patients. Therefore, OH2 cancer treatment requires efficient and convenient predictive factors to promptly and accurately identify responders, optimize treatment plans, improve treatment outcomes, and avoid or reduce unnecessary economic burdens on non-responders. Summary of the Invention

[0008] In view of this, the purpose of this disclosure is to provide the application of neutrophil count in predicting the efficacy of oncolytic virus therapy for tumors, and a method for treating tumor patients in need based on this efficacy prediction. The embodiments of this disclosure, by testing the neutrophil count level in tumor patients, not only help to more accurately screen tumor patient groups suitable for OH2 therapy, but also provide a scientific basis for optimizing OH2 treatment regimens and improving the efficacy of OH2 therapy.

[0009] The first aspect of this disclosure provides a method for treating tumors using an oncolytic virus, comprising: determining the level of neutrophils in the blood of a tumor patient before tumor treatment; and administering the oncolytic virus to the tumor patient for treatment based on the neutrophil level in the blood of the tumor patient being below a cutoff value, wherein the cutoff value is 2 × 10⁻⁶. 9 Up to 5.5×10 9 Any value in / L.

[0010] In some embodiments, the oncolytic virus is an OH2 oncolytic virus.

[0011] In some embodiments, the oncolytic virus treatment is OH2 oncolytic virus monotherapy.

[0012] In some embodiments, the cutoff value is 3.8 × 10⁻⁶. 9 Up to 4.1×10 9 Any value in / L.

[0013] In some embodiments, the cutoff value is 4.0 × 10⁻⁶. 9 / L.

[0014] In some embodiments, the neutrophil count level is the absolute neutrophil count in the blood.

[0015] In some embodiments, the OH2 oncolytic virus is in the form of an injection solution.

[0016] In some embodiments, the oncolytic virus is delivered via intratumoral injection.

[0017] In some embodiments, the oncolytic virus is delivered by direct subcutaneous injection or by ultrasound-guided injection.

[0018] In some embodiments, the dosage of the OH2 oncolytic virus is determined based on the longest diameter of the tumor, wherein: for tumors with a longest diameter ≤ 1.5 cm, up to 1 mL of the OH2 oncolytic virus is applied; for tumors with a longest diameter < 1.5 cm and a longest diameter ≤ 2.5 cm, up to 2 mL of the OH2 oncolytic virus is applied; for tumors with a longest diameter < 2.5 cm and a longest diameter ≤ 5.0 cm, up to 4 mL of the OH2 oncolytic virus is applied; and for tumors with a longest diameter > 5 cm, up to 8 mL of the OH2 oncolytic virus is applied.

[0019] In some embodiments, the tumor includes at least one of melanoma, colorectal cancer, esophageal cancer, head and neck tumors, gastric cancer, biliary system tumors, and sarcoma.

[0020] In some embodiments, the tumor is a melanoma.

[0021] In some embodiments, the sample used to detect the neutrophil count level is peripheral blood from a cancer patient.

[0022] In some embodiments, the OH2 oncolytic virus strain is named H2d3d4-hGF, classified as Herpes Simplex Virus Type 2, deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 3600 and deposit date of February 3, 2010.

[0023] The second aspect of this disclosure also proposes the application of neutrophil count in predicting the efficacy of oncolytic virus therapy for tumors, wherein the neutrophil count level is the neutrophil count level in the blood of tumor patients before tumor treatment.

[0024] The technical solution provided in this disclosure has at least the following advantages compared with the prior art:

[0025] 1. As verified by the embodiments of this disclosure, the level of neutrophil count in tumor patients before treatment is closely related to the efficacy of tumor treatment. Therefore, the level of neutrophil count can be used as a predictive factor for efficacy to more accurately screen the patient population suitable for OH2 treatment, providing a scientific basis for optimizing treatment plans and improving treatment effects.

[0026] 2. The technical solution provided in this disclosure has a simple detection method, low cost, and good stability, and can become a potential predictive and therapeutic tool for clinicians. Detailed Implementation

[0027] To make the objectives, technical solutions, and advantages of this disclosure clearer, the following detailed description is provided in conjunction with embodiments. It should be understood that the specific embodiments described herein are for illustrative purposes only and are not intended to limit the scope of this disclosure.

[0028] In this publication, the term "T-VEC (Talimogene Laherparepvec)," also known as "Imlygic," refers to a genetically modified herpes simplex virus type 1 (HSV-1). It was the first oncolytic virus drug approved by the U.S. Food and Drug Administration (FDA) for the treatment of melanoma patients who develop cutaneous, subcutaneous, and lymph node lesions after initial surgery. T-VEC, administered intratumorally, selectively replicates in tumor cells, leading to tumor cell rupture and death, while simultaneously releasing GM-CSF (granulocyte-macrophage colony-stimulating factor), a leukocyte growth factor, locally in the tumor tissue, which activates a systemic immune response. The mechanism of action of T-VEC includes inducing tumor tissue lysis and stimulating a systemic anti-tumor immune response. As an oncolytic virus drug, T-VEC still requires further improvement in the treatment of patients with advanced melanoma or PD-1-resistant melanoma.

[0029] In this publication, the term "OH2" or "OH2 virus" refers to "recombinant oncolytic type II herpes simplex virus," which was developed using molecular cloning and DNA (unevaluated) homologous recombination techniques. This involved removing the neurotoxic gene ICP34.5 and the immunosuppressive gene ICP47 from the viral genome and inserting a human hGM-CSF expression cassette into the ICP34.5 region. The nucleotide sequence of the expression cassette is shown in SEQ ID NO: 1. The "OH2 virus" strain is named H2d3d4-hGF, with the proposed classification name: Herpes Simplex Virus Type 2 (Latin name: Herpes Simplex Virus Type 2). The depository is the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, with accession number CGMCC No. 3600 and deposit date of February 3, 2010. In this disclosure, "OH2" or "OH2 virus" refers to an oncolytic virus capable of killing tumor cells, and in the specific context of this disclosure, it should be interpreted as an antitumor drug.

[0030] In this disclosure, the term "injection solution" refers to a sterile solution (including emulsions and suspensions) prepared from a drug for injection into the body, as well as a sterile powder or concentrated solution for reconstitution into a solution or suspension immediately before use. Injection solutions act rapidly and reliably, are unaffected by pH, enzymes, food, etc., have no first-pass effect, and can exert systemic or localized effects. They are suitable for patients who cannot take oral medications or are unable to take oral medications. It should be understood that in this disclosure, "OH2 injection solution" consists of "OH2 virus" particles and a buffer for preserving the "OH2 virus."

[0031] In this publication, the term “OH2 treatment” should be understood as treatment with “OH2 virus” or “OH2 virus preparation”, such as treatment with “OH2 injection”.

[0032] In this publication, the term "RECIST" refers to the Standards for Evaluating the Efficacy of Immunotherapy for Solid Tumors, a set of standards used to assess the effectiveness of anti-tumor treatments. It was first developed in 2000 by the National Cancer Institute of the United States and the National Cancer Institute of Canada (v1.0) and revised in 2009 (v1.1). The RECIST criteria provide definitions of cancer treatment efficacy, including complete response (CR), partial response (PR), stable disease (SD), and disease progression (PD). The term "iRECIST" is a revised guideline for evaluating the efficacy of immunotherapy based on the RECIST 1.1 criteria, specifically for clinical trials of immunotherapy for solid tumors. iRECIST addresses the limitations of RECIST 1.1 in immunotherapy by introducing the concepts of "unconfirmed progression" and "confirmed progression," improving the accuracy of assessing immunotherapy efficacy, extending progression-free survival assessment, and better reflecting patient survival benefits.

[0033] In this publication, the term "OS" is an abbreviation for Overall Survival. In medical research, particularly in oncology, OS is a crucial efficacy assessment indicator, measuring the time from randomization (or treatment initiation) to patient death. OS is the gold standard for evaluating the long-term effects of anti-cancer therapy because it directly reflects the impact of treatment on patient survival.

[0034] In this publication, the term “PFS” is an abbreviation for Progression-Free Survival, which refers to the time from the start of randomization (or the start of treatment in a single-arm trial) to tumor progression or death from any cause (whichever comes first). This metric is very important in oncology clinical trials because it provides an intermediate endpoint for measuring treatment efficacy and can assess treatment effectiveness earlier than overall survival (OS).

[0035] In this publication, the term "iORR" refers to an indicator related to the evaluation of the efficacy of cancer (solid tumors only), and "objective response rate (iORR)" refers to the sum of the proportion of partial response (iPR) and complete response (iCR) achieved by immunotherapy; a higher iORR indicates that more patients achieve tumor shrinkage using this immunotherapy method.

[0036] In this publication, the term "DCR" is an abbreviation for Disease Control Rate, a metric used to assess the effectiveness of cancer treatment. It describes the percentage of patients with advanced cancer who achieve complete remission (CR), partial remission (PR), or stable disease (SD) after treatment intervention. Specifically, the DCR is calculated as: DCR = (CR + PR + SD) / number of evaluable cases, which represents the percentage of evaluable cases that achieve remission (including complete and partial remission) or stable disease after treatment.

[0037] In this publication, the term "mOS" stands for median overall survival, also known as the half-life. It refers to the survival time corresponding to a cumulative survival rate of 0.5, meaning that 50% of patients can survive within this timeframe. Median survival is an efficacy indicator for evaluating the survival of cancer patients. Simply put, median survival refers to the time during which 50% of patients can still survive after a certain treatment. Median survival is often used to assess the prognosis of patients with malignant tumors; a longer median survival time indicates a longer overall survival for patients with malignant tumors. Furthermore, the length of median survival can also indicate the effectiveness of new treatment regimens. If clinical studies find that patients receiving a certain treatment regimen have a prolonged median survival, or a median survival significantly exceeding that of the current standard treatment regimen, then the new treatment regimen can usually be recommended or applied in clinical practice, bringing certain benefits to patients, such as increasing their survival rate and improving their quality of life.

[0038] In this publication, the term "ANC" is an abbreviation for Absolute Neutrophil Count, which refers to the number of neutrophils in the blood. The term "baseline ANC" refers to the number of neutrophils in the blood of a patient or subject prior to receiving a cancer treatment regimen.

[0039] In this publication, the term "cutoff value" refers to a predetermined boundary value (critical value) or threshold used to distinguish between different outcomes (such as normal versus abnormal, negative versus positive). The "cutoff value" is derived based on statistical principles and large sample data analysis to determine whether a medical indicator exceeds the normal range, thereby further determining whether intervention or treatment is necessary.

[0040] In this disclosure, the term "preferred" is used only to describe a more effective implementation or embodiment, and should be understood not to limit the scope of protection of this disclosure.

[0041] Neutrophils are an important type of white blood cell, playing a crucial role in the human immune system. Normal neutrophil counts are essential for assessing an individual's health. However, current research lacks studies on the correlation between neutrophil counts and the efficacy of tumor treatments, and there is no evidence that neutrophil counts are used as a predictor of treatment efficacy or to guide oncolytic virus therapy in the field of oncolytic virus therapy for tumors. This disclosure, disregarding cancer indications, comprehensively analyzes biomarker-related data and efficacy evaluation indicators in a Phase I / II study of OH2 virus for tumor treatment. The aim is to validate the role of neutrophil counts (ANC) as a prognostic factor for melanoma and other cancers, examine its relationship with the clinical response to OH2 virus therapy for tumors, and identify a novel biomarker, ANC, in OH2-treated melanoma and other cancers. The results reveal previously overlooked biological effects of OH2 in tumor treatment and provide support for personalized cancer prognosis and treatment.

[0042] Based on this, embodiments of this disclosure provide the application of neutrophil count in predicting the efficacy of oncolytic virus therapy for tumors, wherein the neutrophil count level is the neutrophil count level in the blood of tumor patients before tumor treatment.

[0043] In some embodiments, the oncolytic virus treatment is OH2 oncolytic virus monotherapy in the application.

[0044] In some embodiments, in the said application, the neutrophil count is used to predict the efficacy of tumor treatment, and the critical value for the neutrophil count used to predict the efficacy of tumor treatment is 2 × 10⁻⁶. 9 ~5.5×10 9 Any value in / L; when cancer patients have a baseline ANC level below the threshold in their blood before receiving treatment, the efficacy of OH2 monotherapy is better than that of cancer patients with a baseline ANC level above the threshold in their blood.

[0045] In some embodiments, in the said application, the neutrophil count is used to predict the efficacy of tumor treatment, and the critical value for the neutrophil count used to predict the efficacy of tumor treatment is 3.8 × 10⁻⁶. 9 ~4.1×10 9 Any value in / L; when cancer patients have a baseline ANC level below the threshold in their blood before receiving treatment, the efficacy of OH2 monotherapy is better than that of cancer patients with a baseline ANC level above the threshold in their blood.

[0046] In some embodiments, in the application of this study, the neutrophil count is used to predict the efficacy of tumor treatment, and the critical value for the neutrophil count used to predict the efficacy of tumor treatment is 4.0 × 10⁻⁶. 9 / L; When the baseline ANC level in the blood of cancer patients is 4.0 × 10⁻⁶ before treatment. 9 Tumor subjects with tumors below / L showed better efficacy with OH2 monotherapy than those with baseline ANC levels above 4.0 × 10⁶. 9 The efficacy of OH2 monotherapy in tumor subjects with a tumor size of / L.

[0047] In some embodiments, the applicable tumor types include at least one of melanoma, colorectal cancer, esophageal cancer, head and neck tumors, gastric cancer, biliary system tumors, and sarcoma.

[0048] In some embodiments, the sample used to detect neutrophil count levels is peripheral blood from a cancer patient.

[0049] This disclosure also provides a method for treating tumors using an oncolytic virus, comprising: determining the level of neutrophils in the blood of a tumor patient before tumor treatment; and administering the oncolytic virus to the tumor patient for treatment based on the neutrophil level in the blood of the tumor patient being below a cutoff value, wherein the cutoff value is 2 × 10⁻⁶. 9 Up to 5.5×10 9 Any value in / L.

[0050] In some embodiments, the oncolytic virus is an OH2 oncolytic virus, and the oncolytic virus treatment is OH2 oncolytic virus monotherapy.

[0051] In some embodiments, the cutoff value is 3.8 × 10⁻⁶. 9 Up to 4.1×10 9 Any value in / L.

[0052] In some embodiments, the cutoff value is 4.0 × 10⁻⁶. 9 / L.

[0053] In some embodiments, the oncolytic virus is delivered via intratumoral injection.

[0054] In some embodiments, the oncolytic virus is delivered by direct subcutaneous injection or by ultrasound-guided injection.

[0055] In some embodiments, the tumor is melanoma.

[0056] Example

[0057] This disclosure provides a clinical trial demonstrating the use of neutrophil count levels in predicting the efficacy of OH2 monotherapy for tumors. This clinical trial prospectively analyzed data from patients receiving OH2 monotherapy. The clinical trial was approved by an independent committee of the clinical institution, and all participating subjects signed informed consent forms before enrollment.

[0058] The entire clinical trial established two subject groups for immunotherapy: Subject Group 1: Subjects receiving OH2 monotherapy (N1=44, melanoma); Subject Group 2: Subjects receiving OH2 monotherapy (N2=71, other cancer types besides melanoma). From the clinical data of Subject Group 1, biomarkers and detection thresholds for predicting the efficacy of OH2 therapy for tumors were screened. The second step involved further validation of the screened biomarkers and detection thresholds using clinical data. The specific details of the clinical trial are as follows.

[0059] Clinical Trial Materials and Methods

[0060] 1. Inclusion and exclusion criteria for subjects

[0061] 1.1 Selection Criteria

[0062] (1) Age 18–75 years (inclusive), male or female; (2) Patients with a confirmed diagnosis of malignant tumors by pathology or cytology, such as digestive system tumors, head and neck tumors, soft tissue sarcomas, melanoma, etc.; (3) Patients with advanced (unresectable or recurrent / metastatic) disease who have failed standard treatment (disease progression or intolerance) or who lack effective treatment options; (4) General condition score ECOG 0-1 point; (5) Expected survival of more than 3 months; (6) Previous anti-tumor treatment (including endocrine, chemotherapy / radiotherapy, targeted therapy) ended more than 4 weeks ago (those who used nitrosoureas and mitomycin chemotherapy stopped for more than 6 weeks), and have recovered from the adverse reactions of previous treatment to grade 1; (7) Those who have undergone major surgery need to be more than 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) Asymptomatic central nervous system metastases, or asymptomatic brain metastases after treatment, must be shown no disease progression by computed tomography (CT) or magnetic resonance imaging (MRI), stable for at least 3 months, and have not needed steroid drug treatment for at least 4 weeks; (11) Laboratory test: WBC ≥ 3.0 × 10 9 / L, ANC≥1.5×10 9 / L, PLT≥100×10 9 / L, Hb≥90g / L; serum 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 metastases not exceeding 5 times the upper limit of normal; normal coagulation function (PT, APTT within 1.5 times the upper limit of normal); (12) female subjects and their spouses use effective contraception during treatment and for 3 months after treatment; (13) subjects with herpes need to be treated for 3 months after the end of herpes; (14) voluntarily sign informed consent form, and expected compliance is good.

[0063] 1.2 Exclusion criteria:

[0064] (1) Concurrent serious medical conditions, including severe heart disease, cerebrovascular disease, uncontrolled diabetes, uncontrolled hypertension, severe infection, or active peptic ulcer; (2) Symptomatic brain metastases; (3) Active infection or unexplained fever >38.5°C during screening or before first administration; (4) Congenital or acquired immunodeficiency (e.g., HIV infection) or active hepatitis (hepatitis B reference: HBsAg, Anti-HBs, HBeAg, Anti-HBc, Anti-HBe, HBV DNA (not evaluated) ≥10). 4 / mL, hepatocyte transaminase, etc.; Hepatitis C reference: HCV antibody and HCV RNA (not evaluated), etc.); (5) The lesion cannot meet the requirement of 1mL volume for intratumoral injection; (6) Pregnant or lactating women; (7) Other experimental drug treatment or antiviral treatment used or currently used within 4 weeks before treatment; (8) Other clinical studies participated in within the past 4 weeks; (9) Patients allergic to herpes virus and drug components; (10) Subjects have a known history of psychotropic drug abuse, alcoholism or drug use; (11) Those who have suffered from other malignant tumors within 5 years before enrollment, excluding cervical carcinoma in situ that has been effectively resected, low-risk gastrointestinal stromal tumors, breast cancer, basal cell carcinoma of the skin, squamous cell carcinoma of the skin, 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 by hormone replacement therapy alone), c. Controlled breast cancer d. Skin diseases that do not require systemic treatment (e.g., vitiligo, psoriasis, alopecia), e. any other disease that will not recur without external triggering factors; (13) patients who have been on systemic treatment with corticosteroids (prednisone >10 mg / day or equivalent dose of the same drug) or other immunosuppressants within 14 days prior to administration of the study drug, but may be enrolled if they have currently or previously used any of the following steroid regimens: a. adrenaline replacement steroids (prednisone ≤10 mg / day or equivalent dose of the same drug), b. topical, ophthalmic, intra-articular, intranasal or inhaled corticosteroids with very low 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 to contact allergens); (14) the investigator considers the patient unsuitable for participation in this trial for any reason.

[0065] 2. Treatment and Assessment Methods

[0066] 2.1 Treatment Method: OH2 is delivered via intratumoral injection, directly to skin or subcutaneous lesions, or under ultrasound guidance for deep lymph node or organ metastases. The volume injected into each lesion is based on the longest diameter of the tumor: ≤1.5cm, maximum 1mL; >1.5 to ≤2.5cm, maximum 2mL; >2.5 to ≤5.0cm, maximum 4mL; >5cm, maximum 8mL. Injection into various tumor tissues is permitted. There is no limit to the number of lesions that can be injected per patient, but the maximum volume that can be injected during each visit is 8mL.

[0067] 2.2 Efficacy assessment: Clinical efficacy of solid tumors was assessed according to the iRECIST criteria.

[0068] 3. Statistical Analysis Methods

[0069] Baseline characteristics of patients were described using descriptive statistical methods. As needed, χ² was used to analyze these characteristics. 2- The correlation between ANC and patient characteristics was analyzed using the Mann-Whitney U test. A comprehensive survival analysis was performed on overall survival (OS) and progression-free survival (PFS). Survival analyses employed the Kaplan-Meier method and log-rank test. A univariate Cox proportional hazards regression model was further applied to identify independent prognostic factors for ANC, adjusting for potential confounding variables such as age, sex, ECOG score, number of prior lines of treatment, clinical stage, and duration of OH2 treatment. Results are presented as hazard ratios (HR) and 95% confidence intervals (95% CI).

[0070] 4. Results and Analysis

[0071] This disclosure further analyzes the relationship between the treatment efficacy and absolute neutrophil count (ANC) in each group of patients. The ANC value is the baseline ANC, which is the neutrophil count in the blood of each subject before receiving the treatment regimen.

[0072] Table 1 shows the correlation between baseline ANC levels and best overall efficacy (BOR) in patients in the first experimental group (melanoma indication Ia / Ib, n=44) after receiving OH2 monotherapy.

[0073] Table 1

[0074] As shown in Table 1, the best overall response for the 10 subjects was partial response (PR), with a mean active disease response (ANC) of 3.1 × 10⁻⁶. 9 / L, the best overall efficacy among 14 subjects was SD, and the average ANC was 3.6×10. 9 / L, the best overall efficacy among 18 subjects was PD, and the average ANC was 4.0×10 9 These results indicate that the ANC level in melanoma patients is negatively correlated with the clinical efficacy of OH2 monotherapy, meaning that subjects with low ANC values ​​benefit more from OH2 monotherapy than those with high ANC values.

[0075] To define the population with high ANC, the third quartile of the ANC value among the 44 subjects in the first experimental group was selected as the absolute cutoff value (4.44 × 10⁻⁶). 9The values ​​were used for further univariate grouping analysis. Table 2 shows the results of univariate grouping analysis of iPFS and iORR in the first treatment group (OH2 monotherapy), where iPFS (HR = 0.3; 95% CI 0.1-0.7; P = 0.007) and iORR (OR = 8.5; 95% CI 0.5-159.8; P = 0.100) indicate that the low ANC population (ANC ≤ 4.4) has a better prognosis and a higher response rate. OH2 monotherapy in the low ANC population significantly enriched patients with better sustained partial remission (PR) and stable disease (SD), and both the objective response rate (iORR) and disease control rate (DCR) were significantly improved.

[0076] Table 2

[0077] To determine the optimal cutoff point for baseline ANC, this embodiment of the disclosure used different baseline ANC cutoff values ​​(2.0 × 10⁻⁶) for subjects in the first experimental group. 9 / L, 2.5×10 9 / L, 3.0×10 9 / L, 3.5×10 9 / L, 4.0×10 9 / L, 4.5×10 9 / L, 5.0×10 9 / L, 5.5×10 9 A univariate grouping analysis was performed on / L), and the results are shown in Table 3.

[0078] Table 3

[0079] As can be seen from the results in Table 3, the baseline ANC critical values ​​are 2.0 × 10⁻⁶. 9 / L, 2.5×10 9 / L, 3.0×10 9 / L, 3.5×10 9 / L, 4.0×10 9 / L, 4.5×10 9 / L, 5.0×10 9 / L, 5.5×10 9 At a concentration of 2.0 × 10⁹ / L, tumor patients with baseline ANC levels below each threshold showed significantly better efficacy with OH₂ monotherapy than tumor patients with baseline ANC levels above each threshold. This indicates that the baseline ANC threshold can be selected from 2.0 × 10⁹ / L. 9 ~5.5×10 9 Any value in / L. The results in Table 3 also show that only at 4.0 × 10 9At a cutoff value of 4.0 × 10⁹ / L, the difference between PFS and iORR was statistically significant. 9 / L can be used as the optimal critical value for baseline ANC level.

[0080] To explore the optimal cutoff range for baseline ANC levels, based on the baseline ANC levels and efficacy data of the 44 subjects in the first experimental group, a further threshold of 3.6 × 10⁻⁶ was set. 9 / L, 3.7×10 9 / L, 3.8×10 9 / L, 3.9×10 9 / L, 4.1×10 9 / L, 4.2×10 9 / L, 4.3×10 9 / L, 4.4×10 9 The optimal critical values ​​of eight baseline ANC levels, including / L, were analyzed using univariate grouping analysis. The results are shown in Table 4 below.

[0081] Table 4

[0082] As can be seen from the results in Table 4, the baseline ANC is only 3.8 × 10⁻⁶. 9 / L, 3.9×10 9 / L, 4.0×10 9 / L, 4.1×10 9 When / L is used as the cutoff value, the difference between miPFS and iORR is statistically significant. Therefore, 3.8 × 10⁻⁶ can be selected. 9 ~4.1×10 9 Any value in / L is used as the preferred baseline ANC threshold.

[0083] Take 4.0 × 10 9 / L is the ANC cutoff value. The 44 subjects in the first experimental group were divided into a high ANC group (ANC>4.0×10). 9 / L) and low ANC group (ANC≤4.0×10 9 The miPFS, iORR, and DCR of this group of subjects were analyzed, and the results are shown in Table 5.

[0084] Table 5

[0085] As can be seen from the results in Table 5, the low ANC group (ANC≤4.0×10⁻⁶) in the first test group... 9 In the cohort of subjects with an average annual disease rate (ANC) of 34.5%, a disease control rate (DCR) of 69%, and a median progression-free survival (miPFS) of 3.7 months; in contrast, the high ANC group (ANC>4.0×10⁻⁶) in the first treatment group...9 In the cohort of subjects with ANC (ANC > 4), the iORR (objective response rate) was 0, the DCR (disease control rate) was 38.5%, and the miPFS (median progression-free survival) was 1.8 months. miPFS (HR = 2.9; 95% CI 1.4–6.1), iORR (OR = 0.1; 95% CI 0.0–1.3), and DCR (OR = 0.3; 95% CI 0.1–1.1) indicate that the high ANC population (ANC > 4) has a worse prognosis and lower response rate. These results suggest that setting the baseline ANC cutoff value at 4.0 × 10⁻⁶ is appropriate. 9 When the baseline ANC is 4.0 × 10⁹ / L, it can more accurately predict the optimal efficacy of OH2 monotherapy in melanoma patients, i.e., baseline ANC > 4.0 × 10⁹ / L. 9 At a concentration of 99.9 g / L, patients receiving OH2 monotherapy had a lower response rate, with a baseline ANC ≤ 4.0 × 10⁹ / L. 9 At a concentration of 99.9 g / L, patients receiving OH2 monotherapy had a higher response rate, with a baseline ANC ≤ 4.0 × 10⁹ / L. 9 Patients with melanoma of / L are more likely to benefit from OH2 monotherapy.

[0086] From November 2018 to March 2023, 44 melanoma patients in the first treatment group were enrolled in the clinical trial (NCT04386967) and received OH2 monotherapy. Among them, 15 patients had a baseline ANC > 4.0 × 10⁻⁶. 9 / L, and the baseline ANC of the other 29 patients was ≤4.0×10 9 / L, detailed patient demographic characteristics are shown in Table 6.

[0087] In addition, 71 patients with non-melanoma solid tumors (second subject group) who received OH2 monotherapy from a multi-cohort, multi-indication Phase I / II clinical trial (NCT03866525) were also included in the analysis. Most of these patients (n=30) had colorectal cancer, with 30 patients having a baseline ANC > 4.0 × 10⁻⁶. 9 / L, and the baseline ANC of 41 other patients was ≤4.0×10 9 / L, detailed patient demographic characteristics are shown in Table 6. Data showed that baseline factors such as age, sex, disease, and disease stage did not reach statistical significance (all P>0.05), and there was no significant difference between patients with high and low baseline ANC levels.

[0088] Table 6

[0089] This disclosure analyzed the baseline ANC of 71 non-melanoma tumor patients in the second test group described above, and the baseline ANC cutoff value was determined to be 4.0 × 10⁻⁶.9 The mOS, miPFS, iORR, and DCR of the patients were statistically analyzed, as shown in Table 7.

[0090] Table 7

[0091] As can be seen from the results in Table 7, the low ANC group (ANC≤4.0×10⁻⁶) in the second test group... 9 In the cohort of subjects with a mean overall survival (mOS) of 10.1 months, a mean progression-free survival (miPFS) of 1.4 months, an inverse objective response rate (iORR) of 5.7%, and a disease control rate (DCR) of 34.3%; in contrast, the high active disease response rate (ANC) group in the second treatment group (ANC > 4.0 × 10⁻⁶ L) showed significantly better results. 9 In the cohort of subjects with a baseline ANC of 4.0 × 10⁹ / L, the median overall survival (mOS) was 4.2 months, the miPFS was 1.4 months, the inverse ORR was 4.0%, and the disease control rate (DCR) was 20%. The results (mOS = 2.6; 95% CI 1.5–4.4), miPFS (HR = 1.4; 95% CI 0.8–2.2), iORR (OR = 0.7; 95% CI 0.1–10.0), and DCR (OR = 0.5; 95% CI 0.1–1.7) indicate that a high ANC (ANC > 4) population has a worse prognosis and lower response rate. These results suggest that setting the baseline ANC cutoff value at 4.0 × 10⁹ / L is appropriate. 9 When the baseline ANC is 4.0 × 10⁹ / L, it can more accurately predict the optimal efficacy of OH2 monotherapy in patients with tumors other than melanoma, i.e., baseline ANC > 4.0 × 10⁹ / L. 9 At a concentration of 99.9 g / L, patients receiving OH2 monotherapy had a lower response rate, with a baseline ANC ≤ 4.0 × 10⁹ / L. 9 At a concentration of 99.9 g / L, patients receiving OH2 monotherapy had a higher response rate, with a baseline ANC ≤ 4.0 × 10⁹ / L. 9 Patients with melanoma of / L are more likely to benefit from OH2 monotherapy.

[0092] In summary, neutrophil levels, as a key indicator of systemic inflammatory response, are closely related to tumor development and prognosis. They can serve as a biomarker for predicting the efficacy of OH2 therapy in cancer patients, with a predictive threshold of 2 × 10⁻⁶. 9 ~5.5×10 9 For any value in / L, i.e., a baseline ANC level in the blood below the threshold, the efficacy of OH2 treatment is significantly better in tumor subjects with a baseline ANC level above the threshold than in tumor subjects with a baseline ANC level above the threshold. Among these threshold values, the preferred baseline ANC level range is 3.8 × 10⁻⁶. 9 ~4.1×10 9 / L, the optimal baseline ANC level threshold is 4×10 9 / L. Additionally, the baseline ANC level is 4×10 9 Patients with tumors having low ANC levels (below / L) are recommended to receive OH2 monotherapy, which has shown significant efficacy.

[0093] The present disclosure has been described in detail above. Specific examples have been used to illustrate the principles and implementation methods of the present disclosure. The descriptions of the embodiments above are only for the purpose of helping to understand the present disclosure and its core ideas. It should be noted that those skilled in the art can make several improvements and modifications to the present disclosure without departing from the principles of the present disclosure, and these improvements and modifications also fall within the protection scope of the claims of the present disclosure.

Claims

1. A method for treating tumors using oncolytic viruses, comprising: Determine the level of neutrophils in the blood of cancer patients before cancer treatment; and The oncolytic virus was administered to the tumor patient based on a neutrophil count in the blood below a cutoff value, wherein the cutoff value was 2 × 10⁻⁶. 9 Up to 5.5×10 9 Any value in / L.

2. The method according to claim 1, wherein the oncolytic virus is an OH2 oncolytic virus, and the oncolytic virus treatment is OH2 oncolytic virus monotherapy.

3. The method according to claim 1, wherein the cutoff value is 3.8 × 10⁻⁶. 9 Up to 4.1×10 9 Any value in / L.

4. The method according to claim 1, wherein the cutoff value is 4.0 × 10⁻⁶. 9 / L.

5. The method of claim 1, wherein the neutrophil count level is the absolute neutrophil count in the blood.

6. The method according to claim 2, wherein the OH2 oncolytic virus is in the form of an injection solution.

7. The method of claim 1, wherein the oncolytic virus is delivered via intratumoral injection.

8. The method of claim 1, wherein the oncolytic virus is delivered by direct subcutaneous injection or by ultrasound-guided injection.

9. The method of claim 8, wherein the amount of the OH2 oncolytic virus is determined based on the longest diameter of the tumor, wherein: Based on the tumor's longest diameter being ≤1.5cm, a maximum of 1mL of the OH2 oncolytic virus is administered; Based on the condition that the longest diameter of the tumor is less than 2.5 cm and the maximum amount of the OH2 oncolytic virus is 1.5 cm, a maximum of 2 mL of the virus may be administered. Based on the condition that the longest diameter of the tumor is ≤5.0 cm and 2.5 < the maximum diameter of the tumor, a maximum of 4 mL of the OH2 oncolytic virus may be administered. Based on the tumor's longest diameter >5cm, administer up to 8mL of the OH2 oncolytic virus.

10. The method according to claim 1, wherein the tumor includes at least one of melanoma, colorectal cancer, esophageal cancer, head and neck tumors, gastric cancer, biliary system tumors, and sarcoma.

11. The method according to claim 1, wherein the tumor is a melanoma.

12. The method of claim 1, wherein the sample used to detect the neutrophil count level is peripheral blood from a cancer patient.

13. The method according to claim 2, wherein the OH2 oncolytic virus strain is named H2d3d4-hGF, classified as Herpes Simplex Virus Type 2, deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 3600 and deposit date of February 3, 2010.

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