Marker for predicting or evaluating efficacy of Anti-il-4rα monoclonal antibody in treatment of type-2-inflammation-related nasal disease in nasal mucosa, and method and use

Abstract

Disclosed in the present invention is a marker for predicting or evaluating the efficacy of an anti-IL-4Rα monoclonal antibody in the treatment of type-2-inflammation-related nasal disease in nasal mucosa. The marker is any one of TARC, TNF-α, TGF-β2, 15[S]-HETE, IL-25, CST1, CLC and ALOX15. Optionally, the marker can also be used for predicting or evaluating the degree of inflammation of type-2-inflammation-related nasal disease in nasal mucosa. The marker of the present invention can quickly and accurately predict or evaluate the efficacy of an anti-IL-4Rα monoclonal antibody in the treatment of type-2-inflammation-related nasal disease in nasal mucosa.

Description

A marker, method and application for predicting or evaluating the efficacy of anti-IL-4Rα monoclonal antibody in treating nasal diseases associated with type 2 inflammation of the nasal mucosa Technical Field

[0001] The present invention relates to the field of disease analysis technology, and specifically to a marker, method and application for predicting or evaluating the efficacy of anti-IL-4Rα monoclonal antibodies in treating nasal diseases associated with type 2 inflammation of the nasal mucosa. Background Art

[0002] Type 2 inflammation is characterized by massive tissue eosinophil infiltration, excessive immunoglobulin E (IgE) secretion, and elevated expression of interleukins (IL)-4, IL-5, and IL-13. Respiratory diseases such as chronic rhinosinusitis with nasal polyps (CRSwNP), allergic rhinitis (AR), and asthma all have type 2 inflammation as a major pathogenesis. Anti-IL-4 receptor α subunit (IL-4Rα) monoclonal antibodies (e.g., dupilumab or CM310) are monoclonal antibodies targeting IL-4Rα. They work by blocking the binding of IL-4 and IL-13, key drivers of type 2 inflammatory signaling, to IL-4Rα, inhibiting its biological activity.

[0003] Dupilumab has been approved by the US Food and Drug Administration and the European Medicines Agency for the treatment of CRSwNP due to its excellent efficacy and safety. However, it is expensive, with annual treatment costs exceeding US$50,000. However, a Phase III clinical trial of dupilumab demonstrated that only 62%-65% of CRSwNP patients benefited from 24 weeks of treatment. CM310, the first domestically produced anti-IL-4Rα monoclonal antibody, has demonstrated efficacy and safety in a Phase II clinical trial. Although all patients in our study had eosinophilic CRSwNP, 21% of patients did not benefit from treatment.

[0004] In the field of AR, there is a lack of clinical trials investigating the efficacy and safety of anti-IL-4Rα monoclonal antibodies. A post-hoc analysis of only one Phase II asthma clinical study showed that dupilumab could improve AR-related nasal symptoms and patients' quality of life. Therefore, we conducted the world's first clinical trial of an anti-IL-4Rα monoclonal antibody (CM310) for the treatment of seasonal AR (SAR). We found that CM310 monoclonal antibody significantly improved nasal symptoms, ocular symptoms, and quality of life in subjects with peripheral blood eosinophil counts ≥300 cells / μL. However, among the overall subjects, CM310 monoclonal antibody only showed a trend toward improving the clinical symptoms of SAR patients and had no significant advantage over placebo (mometasone furoate nasal spray + loratadine).

[0005] This suggests that monoclonal antibodies are expensive and their effectiveness in treating type 2 nasal inflammation-related nasal diseases is population-specific. Currently, there are no local nasal mucosal biomarkers that predict monoclonal antibody efficacy. Therefore, screening for such biomarkers to identify sensitive populations will help reduce the economic burden and improve treatment efficacy. Summary of the Invention

[0006] The present invention first provides a marker for predicting or evaluating the efficacy of anti-IL-4Rα monoclonal antibody in treating nasal diseases related to type 2 inflammation of the nasal mucosa, wherein the marker is any one of TARC, TNF-α, TGF-β2, 15[S]-HETE, IL-25, CST1, CLC, and ALOX15;

[0007] Optionally, the markers can also predict or assess the degree of inflammation of nasal diseases associated with type 2 inflammation of the nasal mucosa.

[0008] Preferably, the marker is ALOX15.

[0009] The present invention also provides applications of the above-mentioned markers, characterized in that the applications are one or more of the following:

[0010] a) Prepare a test kit to predict or evaluate the efficacy of anti-IL-4Rα monoclonal antibodies in treating nasal diseases associated with type 2 inflammatory bowel disease;

[0011] b) Prepare or evaluate a test kit for predicting or evaluating the efficacy of anti-IL-4Rα monoclonal antibodies in treating nasal diseases associated with type 2 inflammation of the nasal mucosa;

[0012] c) a method for predicting or evaluating the efficacy of anti-IL-4Rα monoclonal antibodies in treating nasal diseases associated with type 2 inflammation of the nasal mucosa,

[0013] d) preparing a detection reagent for predicting or evaluating the degree of inflammation of nasal diseases associated with type 2 inflammation of the nasal mucosa;

[0014] e) preparing or evaluating a test kit for predicting or evaluating the degree of inflammation of nasal diseases associated with type 2 inflammation of the nasal mucosa;

[0015] f) Methods for predicting or assessing the degree of inflammation in nasal diseases associated with type 2 inflammation of the nasal mucosa;

[0016] The methods are optionally used for non-diagnostic or non-therapeutic purposes.

[0017] The present invention also provides a kit for predicting or evaluating the efficacy of anti-IL-4Rα monoclonal antibodies in treating nasal diseases associated with type 2 inflammation of the nasal mucosa, the kit comprising a detection reagent for detecting the above-mentioned marker;

[0018] Optionally, the kit can also predict or assess the degree of inflammation of nasal diseases associated with type 2 inflammation of the nasal mucosa.

[0019] The present invention also provides the use of the above-mentioned kit in the preparation of a method for predicting or evaluating the efficacy of anti-IL-4Rα monoclonal antibodies in treating nasal diseases associated with type 2 inflammation of the nasal mucosa or the degree of inflammation of nasal diseases associated with type 2 inflammation of the nasal mucosa, wherein the use is optionally used for non-diagnostic or non-therapeutic purposes.

[0020] The present invention also provides the use of a detection reagent for the above-mentioned marker in the preparation of a kit for predicting or evaluating the efficacy of an anti-IL-4Rα monoclonal antibody in treating nasal diseases associated with type 2 inflammation of the nasal mucosa or the degree of inflammation of nasal diseases associated with type 2 inflammation of the nasal mucosa.

[0021] The present invention also provides a method for predicting or evaluating the efficacy of an anti-IL-4Rα monoclonal antibody in treating nasal diseases associated with type 2 inflammation of the nasal mucosa. The method is optionally used for non-diagnostic or non-therapeutic purposes, and comprises the steps of detecting ALOX15 expression, and predicting or evaluating the efficacy of the anti-IL-4Rα monoclonal antibody based on the ALOX15 expression result.

[0022] Optionally, the ALOX15 expression level is the ALOX15 mRNA expression level;

[0023] Optionally, the monoclonal antibody is CM310.

[0024] Preferably, the treatment effect of the patients with high ALOX15 expression is better than that of the patients with low ALOX15 expression.

[0025] Preferably, the lower standard for patients with high ALOX15 expression is ALOX15 mRNA ≥ median 0.803, and the lower standard for patients with low ALOX15 expression is ALOX15 mRNA < median 0.803.

[0026] The present invention also provides a method for predicting or assessing the degree of inflammation of a nasal disease associated with type 2 inflammation of the nasal mucosa. The method is optionally used for non-diagnostic or non-therapeutic purposes, and comprises the steps of detecting the expression level of ALOX15, and predicting or assessing the degree of inflammation of the nasal disease associated with type 2 inflammation of the nasal mucosa based on the ALOX15 expression result.

[0027] Optionally, the ALOX15 expression level is the ALOX15 mRNA expression level.

[0028] Compared with the prior art, the present invention has at least the following beneficial effects:

[0029] The present invention discovered that ALOX15 can be used as a marker to predict or evaluate the efficacy of anti-IL-4Rα monoclonal antibodies in treating nasal diseases associated with type 2 inflammation of the nasal mucosa. The marker of the present invention can quickly and accurately predict or evaluate the efficacy of anti-IL-4Rα monoclonal antibodies in treating nasal diseases associated with type 2 inflammation of the nasal mucosa, filling a gap in the existing technology and having significant clinical and economic value. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 Experimental design process;

[0031] Figure 2. Changes in daily rTNSS scores compared with baseline. (A) Average changes in daily rTNSS scores compared with baseline at 2 and 4 weeks after treatment in the high-dose CM310 group, the low-dose CM310 group, and the placebo group. (B) Changes in daily rTNSS scores compared with baseline at different treatment time points in the high-dose CM310 group, the low-dose CM310 group, and the placebo group.

[0032] Figure 3 Changes in baseline symptom scores at different treatment time points in the CM310 high-dose group, low-dose group, and placebo group;

[0033] Figure 4. Changes in ALOX15 and 15(S)-HETE expression before and after CM310 monoclonal antibody treatment. (A) ALOX15 expression in nasal polyp tissue; (B) 15(S)-HETE expression in nasal secretions. DETAILED DESCRIPTION

[0034] In order to make the technical problems, technical solutions and advantages to be solved by the present invention clearer, a detailed description will be given below with reference to the accompanying drawings and specific embodiments.

[0035] Example 1 Patient data and study design

[0036] This study is a multicenter clinical trial led by Beijing Tongren Hospital affiliated to Capital Medical University from August 17, 2022 to December 28, 2022. A total of 92 patients were included in this study, and all patients had to meet the following inclusion criteria: 1) 18 years old ≤ ≤ 65 years old; 2) SAR that meets the diagnostic criteria of the "Guidelines for the Diagnosis and Treatment of Allergic Rhinitis in China (2022, Revised Edition)", with a history of SAR in the same season for at least 2 years; 3) The subjects have an immunoglobulin E (IgE)-mediated hypersensitivity reaction to at least one pollen allergen in the current environment, which can be confirmed based on the results of the skin prick test or specific IgE test performed by the patient in the previous year (before screening). For skin prick test: A positive reaction is defined as a wheal diameter at least 3 mm larger than the negative control. For specific IgE tests, the diagnosis of hypersensitivity reactions will be made according to the current clinical standards of the research center; 4) The subjects have sufficient pollen exposure during the pollen season: it is expected that during the treatment period, the subjects should be within the pollen season throughout the treatment period and have no travel plans to leave the area for 48 hours or more; 5) The subject's medical history before screening shows that during the pollen season of the same period last year, nasal corticosteroids or other SAR treatment drugs (antihistamines, leukotriene receptor antagonists, etc.) were used, and SAR symptoms were poorly controlled (still had at least one moderate or above SAR symptom); 6) The subject's morning (AM) instantaneous total nasal symptom score (iTNSS) before screening was ≥6 points; at the baseline visit, the subject's AM iTNSS was ≥6 points, and the reflective total nasal symptom score (reflective total nasal symptom score) for the past 6 months was ≥6 points. Patients were required to have an average TNSS score of ≥ 6 (defined as three AM and three PM assessments over the past three 24-hour periods, including the AM assessment at the baseline visit), with nasal congestion ≥ 2 points and any of the three symptoms of runny nose, itchy nose, or sneezing ≥ 2 points. This study was approved by the Ethics Committee of Beijing Tongren Hospital, Capital Medical University. Written informed consent was obtained from patients before participation.

[0037] Patients underwent a baseline visit before the first dose of the trial drug. After a screening / run-in period and review of inclusion / exclusion criteria, those who met the eligibility criteria were randomly assigned in a 1:1:1 ratio, stratified by study center, to the low-dose CM310 group (CM310 600-300 mg QW), the high-dose CM310 group (CM310 600-300 mg Q2W), or the placebo group. Basic patient characteristics are shown in Table 1. Patients in the three groups received treatment for a total of four weeks, during which they continued to receive mometasone furoate aqueous nasal spray (MFNS) and loratadine. The experimental procedure is shown in Figure 1.

[0038] Table 1 Basic characteristics of patients

[0039] Example 2 Clinical Evaluation Indicators

[0040] The AM rTNSS and reflective total ocular symptom score (rTOSS), PM rTNSS and rTOSS, and the morning pre-dose iTNSS and iTOSS were assessed daily at baseline and during treatment. After the 4-week treatment period, TNSS and TOSS scores were discontinued. The Rhinoconjunctivitis Quality of Life Questionnaire (RQLQ) was completed at baseline, 2 weeks, and 4 weeks of treatment. A global assessment of treatment response was performed at baseline and 4 weeks of treatment.

[0041] The TNSS is the sum of the four symptom scores for runny nose, nasal congestion, nasal itching, and sneezing, with each symptom scored on a scale of 0 to 3, with 0 = no symptoms, 1 = mild, 2 = moderate, and 3 = severe. The rTNSS is a rating of the severity of nasal symptoms over the past 12 hours, administered both during the day (AM rTNSS) and at night (PM rTNSS). The AM iTNSS is a rating of the severity of nasal symptoms immediately before medication administration each morning. The individual nasal symptom scores are separate scores for runny nose, nasal congestion, nasal itching, and sneezing, with each symptom scored on a scale of 0 to 3, with 0 = no symptoms, 1 = mild, 2 = moderate, and 3 = severe.

[0042] The TOSS is the sum of the three symptom scores for itching, tearing, and redness, with each symptom scored on a scale of 0 to 3, with 0 = no symptoms, 1 = mild, 2 = moderate, and 3 = severe. The rTOSS is a rating of the severity of eye symptoms over the past 12 hours, administered during the day (AM rTOSS) and at night (PM rTOSS). The AM iTOSS is a rating of the severity of eye symptoms immediately before medication administration each morning. The individual ocular symptom scores are separate scores for itching, tearing, and redness, with each symptom scored on a scale of 0 to 3, with 0 = no symptoms, 1 = mild, 2 = moderate, and 3 = severe.

[0043] The RQLQ is a 28-item, self-administered, disease-specific (AR) quality of life questionnaire used by participants to assess their quality of life over a one-week period. Each question is scored from 0 to 6, with higher scores indicating a greater impact on quality of life. An overall quality of life score is obtained by averaging all item scores. Furthermore, the following four domains are defined based on the RQLQ: Sleep domain, consisting of three items from the Sleep module; Daily Life domain, consisting of seven items from the Non-Nose / Ocular Symptoms module and three items from the Practical Problems module; Eye Symptom domain, consisting of four items from the Eye Symptoms module; and Affective domain, consisting of four items from the Affective module.

[0044] The overall assessment of treatment response was based on a 7-point categorical scale in which participants rated their perception of change or lack of change in their allergy symptoms at the end of the study. The 7 categories were: much improved, moderately improved, slightly improved, no change, slightly worsened, moderately worsened, and much worsened.

[0045] For daily rTNSS and daily rTOSS, the number of days with no symptoms or mild symptoms was calculated according to the following definitions: no or mild nasal symptoms were defined as nasal congestion score, nasal itching score, runny nose score, and sneezing score all ≤1 point; no or mild eye symptoms were defined as eye redness score, eye itching score, and tearing score all ≤1 point.

[0046] The onset time was defined as the time when the change from baseline in iTNSS (4, 6, 8, 10, and 12 hours after the first dose) was the first significant decrease (P value < 0.05) in the CM310 group compared with the placebo group.

[0047] The time to peak efficacy was defined as the earliest date when the CM310 group had the largest decrease compared with the placebo group, using the daily rTNSS change from baseline as the reference indicator.

[0048] Statistical methods:

[0049] Mean change from baseline in rTNSS at week 2: An analysis of covariance (ANCOVA) model was used to analyze the mean change from baseline in rTNSS at week 2 as the dependent variable, with baseline rTNSS, center, and treatment group as covariates. Based on the model, the least squares mean of the mean change from baseline in daily rTNSS at week 2 for each group and the difference between the two groups were calculated, along with 97.5% or 95% confidence intervals (for subgroup analyses). P values ​​for testing differences between the two groups were also calculated. Similar ANCOVA models were used to analyze the remaining TNSS-related indicators, nasal single symptom scores, TOSS-related indicators, and eye single symptom scores compared with the baseline mean changes and change rates during the 2nd week of treatment and during the treatment period, the RQLQ total score and each domain score compared with the baseline during the treatment period, and the areas under the curves of the mean changes in daily rTNSS compared with the baseline during the 2nd week of treatment and the entire treatment period. The least squares means and 95% confidence intervals of the mean changes in daily rTNSS compared with the baseline during the 2nd week of treatment and the differences between the two groups were calculated. The P values ​​for testing the differences between the two groups were also calculated.

[0050] Onset of efficacy was analyzed using a mixed-effect model for repeated measures (MMRM) to analyze the change from baseline in iTNSS at each time point (4, 6, 8, 10, and 12 hours after the first dose). The model used the change from baseline at each time point after treatment as the dependent variable, baseline iTNSS as the covariate, and treatment group, center, time, and the interaction between treatment group and time as fixed effects. Based on the model, the least squares means and 95% confidence intervals of the change from baseline at each time point after treatment and the differences between groups were calculated. P values ​​for testing differences between groups were also calculated.

[0051] Descriptive statistics were performed on the number of days with no symptoms or mild symptoms according to daily rTNSS during the 2-week treatment period. One-way analysis of variance was used to calculate the mean number of days with no symptoms or mild symptoms in each group during the 2-week treatment period, as well as the means and 95% confidence intervals of the differences between the two groups. The P values ​​for testing the differences between the two groups were also calculated.

[0052] SAS version 9.4 was used for statistical analysis.

[0053] Example 3 Sample Detection

[0054] Peripheral blood, nasal secretions, and nasal exfoliated cell samples were collected at baseline, week 2, week 4, and week 12 for routine blood tests and histopathological examination for biomarker analysis.

[0055] Peripheral blood specimen collection and testing: Whole blood was collected using procoagulant and anticoagulant blood collection tubes, respectively. The supernatant was collected by centrifugation. The samples were stored in a -80°C freezer until further testing. Serum or plasma cytokine, immunoglobulin, and chemokine concentrations were measured using ELISA or Luminex.

[0056] Obtaining and testing nasal secretion specimens: Place the trimmed expanded sponge into the upper fornix of the left and right nasal cavities to collect nasal secretions and wait for 5 to 10 minutes. After taking it out, place it in the corresponding 15mL centrifuge tubes on the left and right. Dilute the nasal secretions by adding 0.5mL of 0.9% saline and store at 4°C for 2 hours, and obtain the secretion supernatant by centrifugation. The supernatant is divided and stored in a -80°C refrigerator. ELISA and Luminex are used for protein concentration detection later.

[0057] Obtaining and testing nasal exfoliated cell specimens: Take a 15mL centrifuge tube and a nasal swab, brush it repeatedly 5-10 times, remove the nasal swab, and place the nasal swab completely in the 15mL centrifuge tube. After sample preparation is complete, use a 1mL syringe to extract Trizol, add it to two orange 15mL centrifuge tubes and seal them. After sample preparation is completed, it should be stored upright in a -80℃ freezer. Subsequently, RT-qPCR technology will be uniformly used to detect sample mRNA expression.

[0058] Statistical methods:

[0059] The Dwass-Steel-Critchlow-Fligner test was used to compare whether there were any differences in the biomarker change levels between the CM310 treatment group and the placebo group before and after treatment.

[0060] Example 4 Sample Test Results

[0061] After two weeks of treatment with CM310 600-300 mg Q2W, SAR patients showed no significant improvement in rTNSS scores compared with the placebo group, showing only a trend toward improvement. The CM310 600-300 mg Q2W group showed significant improvements compared with baseline in rTNSS and rTOSS scores at week 2, AM iTNSS and AM iTOSS scores at weeks 2 and 4, and nasal congestion scores at weeks 2 and 4, as well as AM iTOSS scores at week 2. No significant improvement in RQLQ scores was observed after CM310 treatment. Changes from baseline in various clinical indicators are shown in Table 2, Figures 1, and 2.

[0062] Table 2 Changes of clinical indicators compared with baseline after CM310 monoclonal antibody treatment

[0063] We measured the following biomarkers in peripheral blood, protein expression in nasal secretions, and mRNA expression in nasal exfoliated cells. CM310 treatment significantly decreased peripheral blood concentrations of thymus and activation-regulated chemokine (TARC), tumor necrosis factor-α (TNF-α), transforming growth factor-β2 (TGF-β2), 15(S)-hydroxyeicosatetraenoic acid (15[S]-HETE), and IgE during treatment and post-treatment follow-up. Protein expression levels of interleukin-25, cystatin-SN (CST1), and Charcot-Leyden crystals (CLC) in nasal secretions were also significantly decreased. The mRNA expression levels of CST1, CLC, and 12 / 15-Lipoxygenase (ALOX15) in nasal exfoliated cells were significantly decreased. The biomarkers whose expression levels changed compared with baseline after CM310 treatment are shown in Table 3.

[0064] Table 3 Biomarker expression levels after CM310 monoclonal antibody treatment

[0065] We divided the subjects into two subgroups based on the median levels of biomarkers in peripheral blood, nasal secretions, and nasal exfoliated cells, namely, the high biomarker group (≥median) and the low biomarker group (<median).

[0066] We analyzed whether there were differences in rTNSS scores between high and low biomarker groups after two weeks of CM310 treatment. We found that, when subjects were stratified by ALOX15 mRNA expression levels in nasal exfoliated cells, subjects with high ALOX15 mRNA expression (ALOX15 mRNA ≥ median [0.803], hereinafter referred to as the high ALOX15 group) showed significant improvement in rTNSS scores at two weeks compared to the control group, regardless of whether they received CM310 600-300 mg QW or CM310 600-300 mg Q2W. However, subjects with low ALOX15 mRNA expression (ALOX15 mRNA < median [0.803], hereinafter referred to as the low ALOX15 group) showed no significant improvement in rTNSS scores at two weeks after treatment with any dose of CM310. This trend was not observed for the other biomarkers. Table 4 shows the changes in rTNSS scores compared with baseline in each ALOX15 subgroup after 2-week treatment with CM310 monoclonal antibody.

[0067] Table 4 Changes in rTNSS scores compared with baseline in the high ALOX15 group and the low ALOX15 group at week 2

[0068] We further evaluated the improvement in other clinical indicators across the ALOX15 subgroups and found that participants in the high-ALOX15 group who received CM310 showed advantages in terms of improved nasal and ocular symptoms, as well as quality of life. The changes in clinical indicators compared to baseline in each ALOX15 subgroup after CM310 treatment are shown in Table 5.

[0069] Table 5 Changes of clinical indicators compared with baseline in the high ALOX15 group and the low ALOX15 group

[0070] Example 5 Results Re-verification

[0071] Patient data and study design:

[0072] Sixteen patients were enrolled in this study. All patients met the following inclusion criteria: 1) aged 18-70 years; 2) received systemic corticosteroids (SCS) within 2 years before the run-in period, or had contraindications or intolerance to SCS, or had undergone nasal polyp surgery within 6 months before the run-in period; 3) had a nasal polyp score (NPS) of at least 5 (at least 2 points for each nostril); 4) had moderate or severe nasal congestion (0 = no symptoms, 1 = mild, 2 = moderate, 3 = severe), and an average weekly nasal congestion score (NPS) of at least 5 points. The study included 5) patients with a nasal steroid injection for at least 4 weeks before screening and MFNS as background therapy; 6) all patients had a peripheral blood eosinophil percentage ≥ 6.9% (without asthma) or ≥ 3.7% (with asthma); 2) nasal polyp biopsy with an absolute tissue eosinophil count ≥ 55 cells / high-power microscopic field or an eosinophil percentage ≥ 27%.

[0073] Patients were excluded if they: 1) had used an IL-4Rα antagonist within 10 weeks or 5 half-lives (whichever was longer) before randomization, or had used biologic therapy / systemic immunosuppressants within 8 weeks or 5 half-lives (whichever was longer), or had used anti-IgE monoclonal antibodies within 130 days; 2) had undergone nasal polyp surgery within 6 months before the screening / run-in period; 3) had received intermediate-acting and short-acting SCS within 4 weeks before screening, or long-acting SCS within 6 weeks; 4) had severe Hepatic or renal impairment; 5) Received live or attenuated vaccines within 12 weeks before randomization, or planned to receive live or attenuated vaccines during the study; 6) Patients with known or suspected immunosuppression, including but not limited to a history of invasive opportunistic infections (e.g., tuberculosis, histoplasmosis, listeriosis, coccidiosis, pulmonary cysts, aspergillosis), even if the infection has resolved; 7) Pregnant or breastfeeding women, or women who plan to become pregnant during the study; 8) Any other medical or non-medical condition deemed inappropriate for study participation by the researchers. This study was approved by the Ethics Committee of Beijing Tongren Hospital, Capital Medical University. Written informed consent was obtained before patient participation.

[0074] After signing the informed consent form, the patients entered a 4-week screening / run-in period, during which all patients received MFNS nasal spray, 100 μg per nasal cavity per day (total daily dose of 200 μg); then, the patients received anti-IL-4Rα monoclonal antibody treatment for 16 weeks (600 mg subcutaneous injection for the first time, followed by 300 mg subcutaneous injection every 2 weeks).

[0075] Sample testing:

[0076] Nasal secretions and nasal polyps were collected from patients at baseline and 16 weeks of treatment to detect the expression of ALOX15 and 15(S)-HETE in nasal secretions and nasal polyp tissues.

[0077] Nasal polyp tissue collection and ALOX15 expression level detection: Typical nasal polyp lesions were selected, and polyp tissue approximately 3 to 5 mm in diameter was removed using ethmoid sinus forceps or bite forceps. The tissue was then paraffin-embedded, and 4 μm sections were immunohistochemically stained using the primary antibody: Anti-15 Lipoxygenase 1, ab244205, abcam; the secondary antibody: HRP-conjugated. Microscopic examination and image acquisition and analysis were performed. Images were captured using a bright-field light microscope, and semi-quantitative immunohistochemical analysis was performed using ImageJ.

[0078] Collection and testing of 15(S)-HETE in nasal secretions: Place the trimmed expanded sponge into the upper fornix of the left and right nasal cavities to collect nasal secretions and wait for 5 to 10 minutes. After removing the long expanded sponge strips, place them in the corresponding green 15mL centrifuge tubes for the left and right nasal cavities. Dilute the nasal secretions by adding 0.5mL of 0.9% saline and store at 4°C for 2 hours. Centrifuge to obtain the supernatant. Aliquot the supernatant and store it in a -80°C refrigerator for subsequent unified 15(S)-HETE protein concentration testing. Testing: Use the 15(S)-HETE ELISA test kit (#534721, Cayman).

[0079] Sample test results:

[0080] In patients with ECRSwNP, ALOX15 protein expression was assessed by immunohistochemistry, and 15(S)-HETE protein expression was detected by ELISA before and after treatment with the anti-IL-4Rα monoclonal antibody CM310 ( Figure 4 ).

[0081] The expression of ALOX15 and 15(S)-HETE was decreased after treatment with CM310 mAb.

[0082] The above is a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A marker for predicting or evaluating the efficacy of anti-IL-4Rα monoclonal antibody in treating nasal diseases associated with type 2 inflammation of the nasal mucosa, characterized in that: The marker is any one of TARC, TNF-α, TGF-β2, 15[S]-HETE, IL-25, CST1, CLC, and ALOX15; Optionally, the marker can also predict or assess the degree of inflammation of nasal diseases associated with type 2 inflammation of the nasal mucosa.

2. The marker according to claim 1, characterized in that The marker is ALOX15.

3. A use of the marker according to claim 1 or 2, characterized in that: The application is one or more of the following; a) Prepare detection reagents for predicting or evaluating the efficacy of anti-IL-4Rα monoclonal antibodies in treating nasal diseases related to type 2 inflammation of the nasal mucosa; b) Prepare or evaluate a test kit for predicting or evaluating the efficacy of anti-IL-4Rα monoclonal antibodies in treating nasal diseases associated with type 2 inflammation of the nasal mucosa; c) a method for predicting or evaluating the efficacy of anti-IL-4Rα monoclonal antibodies in treating nasal diseases associated with type 2 inflammation of the nasal mucosa, d) preparing a detection reagent for predicting or evaluating the degree of inflammation of nasal diseases associated with type 2 inflammation of the nasal mucosa; e) preparing or evaluating a test kit for predicting or evaluating the degree of inflammation of nasal diseases associated with type 2 inflammation of the nasal mucosa; f) Methods for predicting or assessing the degree of inflammation in nasal diseases associated with type 2 inflammation of the nasal mucosa; The methods are optionally used for non-diagnostic purposes or non-therapeutic purposes.

4. A kit for predicting or evaluating the efficacy of anti-IL-4Rα monoclonal antibody in treating nasal diseases associated with type 2 inflammation of nasal mucosa, characterized in that: The kit comprises a detection reagent for detecting the marker according to claim 1 or 2; Optionally, the kit can also predict or assess the degree of inflammation of nasal diseases associated with type 2 inflammation of the nasal mucosa.

5. A use of the kit according to claim 4 in the preparation of a method for predicting or evaluating the efficacy of anti-IL-4Rα monoclonal antibodies in treating nasal diseases associated with type 2 inflammation of the nasal mucosa or the degree of inflammation of nasal diseases associated with type 2 inflammation of the nasal mucosa, wherein the use is optionally used for non-diagnostic purposes or non-therapeutic purposes.

6. Use of a detection reagent for the marker as claimed in claim 1 or 2 in the preparation of a kit for predicting or evaluating the efficacy of anti-IL-4Rα monoclonal antibodies in treating nasal diseases associated with type 2 inflammation of the nasal mucosa or the degree of inflammation of nasal diseases associated with type 2 inflammation of the nasal mucosa.

7. A method for predicting or evaluating the efficacy of anti-IL-4Rα monoclonal antibody in treating nasal diseases associated with type 2 inflammation of nasal mucosa, characterized in that: The method is optionally used for non-diagnostic or non-therapeutic purposes, and comprises the steps of detecting the expression level of ALOX15, and predicting or evaluating the efficacy of anti-IL-4Rα monoclonal antibody based on the result of ALOX15 expression level; Optionally, the ALOX15 expression level is the ALOX15 mRNA expression level; Optionally, the monoclonal antibody is CM310.

8. The method according to claim 7, characterized in that The treatment effect of the patients with high ALOX15 expression is better than that of the patients with low ALOX15 expression.

9. The method according to claim 8, characterized in that The lower standard for patients with high ALOX15 expression is ALOX15 mRNA ≥ median 0.803, and the standard for patients with low ALOX15 expression is ALOX15 mRNA < median 0.

803.

10. A method for predicting or evaluating the degree of inflammation of nasal diseases associated with type 2 inflammation of the nasal mucosa, characterized in that: The method is optionally used for non-diagnostic or non-therapeutic purposes, and comprises the steps of detecting the expression level of ALOX15, and predicting or evaluating the degree of inflammation of nasal diseases associated with type 2 inflammation of the nasal mucosa based on the result of the expression level of ALOX15; Optionally, the ALOX15 expression level is the ALOX15 mRNA expression level.

Images