A biomarker, 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 nasal mucosa

Abstract

The present invention discloses a biomarker for predicting or evaluating the efficacy of anti-IL-4Rα monoclonal antibody in the treatment of nasal mucosa type 2 inflammation-related nasal diseases, characterized in that the biomarker is any one of TARC, TNF-α, TGF-β2, 15[S]-HETE, IL-25, CST1, CLC, ALOX15; optionally, the biomarker can also predict or evaluate the degree of inflammation of nasal mucosa type 2 inflammation-related nasal diseases. The biomarker of the present invention can quickly and accurately predict or evaluate the efficacy of anti-IL-4Rα monoclonal antibody in the treatment of nasal mucosa type 2 inflammation-related nasal diseases, filling the blank of the prior art and having great clinical value and economic value.

Description

Technical Field

[0001] The present invention relates to the technical field of disease analysis, and in particular, to a marker, method and application for predicting or evaluating the efficacy of anti-IL-4Rα monoclonal antibody in the treatment of nasal mucosal type 2 inflammation-related nasal diseases. Background Art

[0002] Type 2 inflammation is mainly characterized by massive infiltration of eosinophils in tissues, excessive secretion of immunoglobulin E (IgE), and high expression of interleukin (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 one of the main pathogenesis. Anti-interleukin-4 receptor alpha subunit (IL-4Rα) monoclonal antibody (e.g., Dupilumab or CM310) is a monoclonal antibody targeting IL-4Rα, which exerts its effect by blocking the binding of IL-4 and IL-13, the key driving factors of type 2 inflammatory response signal transduction, to IL-4Rα and 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, its price is expensive, with an annual treatment cost of more than $50,000. But the phase III clinical trial of Dupilumab showed that only 62%-65% of CRSwNP patients benefited from the 24-week treatment. As the first domestic anti-IL-4Rα monoclonal antibody, CM310 has proven its efficacy and safety in the phase II clinical trial. Although all the patients included in our study were eosinophilic CRSwNP patients, 21% of the patients still could not benefit from the treatment.

[0004] In the field of AR, there is still a lack of clinical trials on the efficacy and safety of anti-IL-4Rα monoclonal antibody. Only a post hoc analysis of a phase II clinical study of asthma showed that Dupilumab could improve AR-related nasal symptoms and the quality of life of patients. Therefore, we conducted the world's first clinical trial of anti-IL-4Rα monoclonal antibody (CM310 monoclonal antibody) in the treatment of seasonal AR (SAR). We found that CM310 monoclonal antibody could significantly improve the nasal symptoms, eye symptoms, and quality of life of subjects with a peripheral blood eosinophil count ≥ 300 / μL. However, in the overall subjects, CM310 monoclonal antibody only showed a tendency to improve the clinical symptoms of SAR patients, and there was no significant advantage compared with the placebo (mometasone furoate nasal spray + loratadine).

[0005] It can be seen that monoclonal antibodies are expensive and the effective rate of treating nasal diseases related to type 2 nasal mucosa inflammation is population-specific. At present, there is a lack of local nasal mucosa biomarkers for predicting the efficacy of monoclonal antibodies. Therefore, screening such biomarkers to identify sensitive populations will help reduce the economic burden and improve the treatment effective rate. Summary of the Invention

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

[0007] Optionally, the biomarker can also predict or evaluate the degree of inflammation of nasal diseases related to type 2 nasal mucosa inflammation.

[0008] Preferably, the biomarker is ALOX15.

[0009] The present invention also provides the application of the above biomarker, characterized in that the application is one or more of the following;

[0010] a) Preparing a detection reagent for predicting or evaluating the efficacy of anti-IL-4Rα monoclonal antibody in treating nasal diseases related to type 2 nasal mucosa inflammation;

[0011] b) Preparing or evaluating a detection kit for predicting or evaluating the efficacy of anti-IL-4Rα monoclonal antibody in treating nasal diseases related to type 2 nasal mucosa inflammation;

[0012] c) A method for predicting or evaluating the efficacy of anti-IL-4Rα monoclonal antibody in treating nasal diseases related to type 2 nasal mucosa inflammation,

[0013] d) Preparing a detection reagent for predicting or evaluating the degree of inflammation of nasal diseases related to type 2 nasal mucosa inflammation;

[0014] e) Preparing or evaluating a detection kit for predicting or evaluating the degree of inflammation of nasal diseases related to type 2 nasal mucosa inflammation;

[0015] f) A method for predicting or evaluating the degree of inflammation of nasal diseases related to type 2 nasal mucosa inflammation;

[0016] The method is optionally used for non-diagnostic purposes or non-therapeutic purposes.

[0017] The present invention also provides a kit for predicting or evaluating the efficacy of anti-IL-4Rα monoclonal antibody in treating nasal diseases related to type 2 nasal mucosa inflammation, and the kit contains a detection reagent for detecting the above biomarker;

[0018] Optionally, the kit can also predict or evaluate the degree of inflammation of nasal diseases related to 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 medicament for predicting or evaluating the efficacy of anti-IL-4Rα monoclonal antibody in the treatment of nasal diseases related to type 2 inflammation of the nasal mucosa or the degree of inflammation of nasal diseases related to type 2 inflammation of the nasal mucosa, and the use is optionally for non-diagnostic purposes or non-therapeutic purposes.

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

[0021] The present invention also provides a method for predicting or evaluating the efficacy of anti-IL-4Rα monoclonal antibody in the treatment of nasal diseases related to type 2 inflammation of the nasal mucosa, and the method is optionally for non-diagnostic purposes or non-therapeutic purposes, and it includes 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 the expression level of ALOX15;

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

[0023] Optionally, the monoclonal antibody is CM310.

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

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

[0026] The present invention also provides a method for predicting or evaluating the degree of inflammation of nasal diseases related to type 2 inflammation of the nasal mucosa, and the method is optionally for non-diagnostic purposes or non-therapeutic purposes, and it includes the steps of detecting the expression level of ALOX15 and predicting or evaluating the degree of inflammation of nasal diseases related to type 2 inflammation of the nasal mucosa based on the result of the expression level of ALOX15;

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

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

[0029] The present invention discovers that ALOX15 can be used as a biomarker for predicting or evaluating the efficacy of anti-IL-4Rα monoclonal antibody in the treatment of nasal mucosa type 2 inflammation-related nasal diseases. The biomarker of the present invention can quickly and accurately predict or evaluate the efficacy of anti-IL-4Rα monoclonal antibody in the treatment of nasal mucosa type 2 inflammation-related nasal diseases, filling the gap in the prior art and having great clinical value and economic value. Brief Description of the Drawings

[0030] Figure 1 Experimental design process;

[0031] Figure 2 Change in daily rTNSS score compared to baseline. (A) Mean change in daily rTNSS score of the high-dose group, low-dose group, and placebo group of CM310 at 2 weeks and 4 weeks of treatment compared to baseline; (B) Change in daily rTNSS score of the high-dose group, low-dose group, and placebo group of CM310 at different treatment time points compared to baseline;

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

[0033] Figure 4 Change in the expression of ALOX15 and 15(S)-HETE before and after treatment with CM310 monoclonal antibody. (A) Expression of ALOX15 in nasal polyp tissue; (B) Expression of 15(S)-HETE in nasal secretions. Detailed Description of the Invention

[0034] To make the technical problems, technical solutions, and advantages to be solved by the present invention clearer, the following will be described in detail with reference to the drawings and specific embodiments.

[0035] Example 1 Patient Information and Research Design

[0036] This study was a multicenter clinical trial led by Beijing Tongren Hospital, 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 ≤ age ≤ 65 years old; 2) SAR that met the diagnostic criteria of "Chinese Guidelines for the Diagnosis and Treatment of Allergic Rhinitis (2022, Revised Edition)", with a history of seasonal SAR for at least 2 years; 3) The subjects had immunoglobulin E (IgE)-mediated hypersensitivity reactions to at least one pollen allergen in the current environment, which could be confirmed according to the results of skin prick tests or specific IgE tests received by the patients in the previous year (before screening). For skin prick tests: A positive reaction was defined as a wheal diameter at least 3 mm larger than the negative control. For specific IgE tests, the diagnosis of hypersensitivity reactions would be made according to the current clinical specifications of the research center; 4) The subjects had sufficient pollen exposure during the pollen season: It was expected that during the treatment period, the subjects should be in the pollen season throughout, and there was no travel plan to leave the area for 48 hours or more; 5) The medical history of the subjects before screening showed that during the same period of the pollen season last year, the use of nasal corticosteroids or other drugs for the treatment of SAR (antihistamines, leukotriene receptor antagonists, etc.) did not result in good control of SAR symptoms (still having at least one moderate or more severe SAR symptom); 6) The total instantaneous nasal symptom score (iTNSS) of the subjects before screening in the morning (AM) ≥ 6 points; At the baseline visit, the AM iTNSS of the subjects ≥ 6 points, and the average score of the total reflective nasal symptom score (rTNSS) in the past 6 reviews ≥ 6 points (i.e., in the past 3 24-hour time periods, 3 AM evaluations and 3 PM evaluations, including the AM evaluation at the baseline visit), among which nasal congestion ≥ 2 points, and at least one of the three symptoms of runny nose, nasal itching, and sneezing ≥ 2 points. This study was approved by the Ethics Committee of Beijing Tongren Hospital, Capital Medical University. Written informed consent was obtained before the patients participated.

[0037] The patients had a baseline visit before the first administration of the investigational drug. After screening / induction period treatment and rechecking the inclusion / exclusion criteria, the subjects who still met the inclusion criteria were randomly assigned to the CM310 low-dose group (CM310 600 - 300 mg QW), the CM310 high-dose group (CM310 600 - 300 mg Q2W), and the placebo group according to a 1:1:1 ratio with the research center as the stratification factor. The basic characteristics of the patients are shown in Table 1. The three groups of patients received treatment for 4 weeks, and during the treatment period, the patients continued to use mometasone furoate aqueous nasal spray (MFNS) and loratadine for treatment. The experimental process is shown inFigure 1 。

[0038] Table 1 Basic characteristics of the patients

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[0041] Example 2 Clinical evaluation indicators

[0042] AM rTNSS and the total reflective ocular symptom score (rTOSS), PM rTNSS and rTOSS, iTNSS before morning medication, and iTOSS before morning medication were evaluated daily at baseline and during treatment. After 4 weeks of treatment, TNSS and TOSS evaluations and records were no longer performed. The Rhinoconjunctivitis Quality of Life Questionnaire (RQLQ) was filled out at baseline, 2 weeks of treatment, and 4 weeks of treatment. The treatment response was evaluated overall at baseline and 4 weeks of treatment.

[0043] TNSS is the sum of the scores of four symptoms: rhinorrhea, nasal congestion, nasal itching, and sneezing. The score for each symptom ranges from 0 to 3, where 0 = no symptoms, 1 = mild, 2 = moderate, and 3 = severe. rTNSS is the rating of the severity of nasal symptoms in the past 12 hours, performed separately during the day (AM rTNSS) and at night (PM rTNSS). AM iTNSS is the rating of the instantaneous severity of nasal symptoms before morning medication each day. The individual nasal symptom score is the separate score of the four symptoms of rhinorrhea, nasal congestion, nasal itching, and sneezing. The score for each symptom ranges from 0 to 3, where 0 = no symptoms, 1 = mild, 2 = moderate, and 3 = severe.

[0044] TOSS is the sum of the scores of three symptoms: eye itching, tearing, and redness. The score for each symptom ranges from 0 to 3, where 0 = no symptoms, 1 = mild, 2 = moderate, and 3 = severe. rTOSS is the rating of the severity of ocular symptoms in the past 12 hours, performed separately during the day (AM rTOSS) and at night (PM rTOSS). AM iTOSS is the rating of the instantaneous severity of ocular symptoms before morning medication each day. The individual ocular symptom score is the separate score of the three symptoms of eye itching, tearing, and redness. The score for each symptom ranges from 0 to 3, where 0 = no symptoms, 1 = mild, 2 = moderate, and 3 = severe.

[0045] The RQLQ is a 28-item, self-administered, disease-specific (AR) quality of life scale used by subjects to evaluate 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. The overall quality of life score is obtained from the average score of all items. In addition, the following 4 domains are defined based on the RQLQ scale: Sleep domain: consists of 3 items in the sleep module; Daily life domain: consists of 7 items in the non-nose / eye symptom module and 3 items in the practical problems module; Eye symptom domain: consists of 4 items in the eye symptom module; Emotional domain: consists of 4 items in the emotional module.

[0046] The overall assessment of treatment response is based on a 7-point categorical scale, on which subjects rate their perception of the change or lack of change in their allergic symptoms at the end of the study. The 7 categories are: marked improvement, moderate improvement, mild improvement, no change, mild worsening, moderate worsening, and marked worsening.

[0047] For daily rTNSS and daily rTOSS, the number of symptom-free or mild days is calculated as follows: Symptom-free or mild nasal symptoms are defined as having a nasal congestion score, nasal itching score, rhinorrhea score, and sneezing score all ≤ 1; Symptom-free or mild eye symptoms are defined as having a red eye score, eye itching score, and tearing score all ≤ 1.

[0048] The onset time is defined as the time when CM310 group first significantly decreased (P value < 0.05) compared with the placebo group, with the change in iTNSS (at 4, 6, 8, 10, and 12 hours after the first dose) relative to the baseline as the reference index.

[0049] The time to peak efficacy is defined as the earliest date when the CM310 group had the largest decrease compared with the placebo group, with the change in daily rTNSS relative to the baseline as the reference index.

[0050] Statistical methods:

[0051] Mean change in rTNSS at week 2 of treatment compared with baseline: Analysis was performed using an analysis of covariance (ANCOVA) model. The model used the mean change in rTNSS at week 2 of treatment compared with baseline as the dependent variable, and baseline rTNSS, center, and treatment group as covariates. Based on the model, the least-squares means and 97.5% or 95% (used in subgroup analysis) confidence intervals of the mean change in daily rTNSS at week 2 of treatment compared with baseline and the differences between groups were calculated, and the P value for testing the differences between groups was also calculated. The mean change and change rate of the remaining TNSS-related indicators, individual nasal symptom scores, TOSS-related indicators, and individual eye symptom scores at week 2 of treatment and during the treatment period compared with baseline, the mean change in the total RQLQ score and scores of each domain during the treatment period compared with baseline, and the area under the curve of the mean change in daily rTNSS at week 2 and throughout the treatment period compared with baseline were all analyzed using a similar ANCOVA model. The least-squares means and 95% confidence intervals of the mean change in daily rTNSS at week 2 of treatment compared with baseline and the differences between groups were calculated, and the P value for testing the differences between groups was also calculated.

[0052] The onset time was analyzed using a mixed-effect model for repeated measures (MMRM) for the change in iTNSS at each time point (4, 6, 8, 10, and 12 hours after the first dose) compared with baseline. The model used the change in iTNSS at each time point after treatment compared with baseline 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 in iTNSS at each time point after treatment compared with baseline and the differences between groups were calculated, and the P value for testing the differences between groups was also calculated.

[0053] Descriptive statistics were performed on the number of days with no symptoms or mild symptoms of daily rTNSS at week 2 of treatment. One-way ANOVA was used to calculate the mean number of days with no symptoms or mild symptoms at week 2 of treatment for each group, the mean value and 95% confidence interval of the differences between groups, and the P value for testing the differences between groups.

[0054] SAS version 9.4 was used for statistical analysis.

[0055] Sample detection in Example 3

[0056] 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 examinations for biomarker analysis.

[0057] Collection and detection of peripheral blood specimens: Whole blood was collected using coagulation-promoting blood collection tubes and anticoagulant blood collection tubes respectively. The supernatant was obtained by centrifugation. The samples were placed in a -80 °C freezer for unified detection. ELISA and Luminex were used to detect the concentrations of cytokines, immunoglobulins and chemokines in serum or plasma.

[0058] Collection and detection of nasal secretion specimens: The trimmed inflated sponge was placed in the upper fornix of the left and right nasal cavities to collect nasal secretions and left for 5 - 10 minutes. After removal, it was placed in a corresponding 15 mL centrifuge tube on the left and right. Diluted nasal secretions were obtained by adding 0.5 mL of 0.9% normal saline and storing at 4 °C for 2 hours, and the supernatant of the secretions was obtained by centrifugation. The supernatant was aliquoted and stored in a -80 °C refrigerator, and ELISA and Luminex were subsequently used for unified protein concentration detection.

[0059] Collection and detection of nasal exfoliated cell specimens: A 15 mL centrifuge tube and a nasal swab were taken, and the nasal cavity was repeatedly swabbed 5 - 10 times. The nasal swab was taken out and completely placed in a 15 mL centrifuge tube. After sample preparation, 1 mL of Trizol was aspirated using a 1 mL syringe and added to 2 orange 15 mL centrifuge tubes respectively and sealed. After sample preparation, it should be stored upright at -80 °C in a freezer, and RT-qPCR technology was subsequently used for unified detection of sample mRNA expression.

[0060] Statistical methods:

[0061] The Dwass-Steel-Critchlow-Fligner test was used to compare whether there were between-group differences in the changes in biomarker levels between the CM310 treatment group and the placebo group before and after treatment.

[0062] Sample test results of Example 4

[0063] After 2 weeks of treatment with CM310 monoclonal antibody 600 - 300 mg Q2W, the rTNSS score of SAR patients showed no significant improvement compared with the placebo group, only showing a tendency to improve. The CM310 monoclonal antibody 600 - 300 mg Q2W group was significantly improved compared with the baseline in terms of the 2-week rTNSS score and rTOSS score, the change rates of the 2-week and 4-week AM iTNSS score and AM iTOSS score, and the 2-week and 4-week nasal congestion scores, 2-week AM iTOSS score and other indicators. The RQLQ score did not show obvious improvement after CM310 monoclonal antibody treatment. The changes in each clinical index compared with the baseline are shown in Table 2, Figure 1 、 Figure 2 。

[0064] Table 2 Changes in clinical indicators after CM310 monoclonal antibody treatment compared with the baseline

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[0076] We detected the concentrations of the following biomarkers in peripheral blood, the protein expressions of biomarkers in nasal secretions, and the mRNA expression levels of biomarkers in nasal exfoliated cells. The study found that after CM310 treatment, the 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 in peripheral blood decreased significantly during the treatment period or the follow-up period after treatment. The protein expression levels of IL-25, cystatin-SN (CST1), and Charcot-Leyden crystal (CLC) in nasal secretions decreased significantly. The mRNA expression levels of CST1, CLC, and arachidonate-15-lipoxygenase (ALOX15) in nasal exfoliated cells decreased significantly. The biomarkers with changes in expression levels after CM310 monoclonal antibody treatment compared with the baseline are shown in Table 3.

[0077] Table 3 Biomarker expression levels after CM310 monoclonal antibody treatment

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[0083] We divided the subjects into two subgroups according to the median levels of each biomarker in peripheral blood, nasal secretions, and nasal exfoliated cells, namely, the high biomarker group (≥ median) and the low biomarker group (< median).

[0084] We analyzed whether there were differences in the improvement of rTNSS scores between the high biomarker group and the low biomarker group after 2 weeks of treatment with CM310 monoclonal antibody. It was found that when the subjects were stratified according to the expression level of ALOX15 mRNA in nasal exfoliated cells, the subjects with high expression of ALOX15 mRNA (ALOX15 mRNA ≥ median [0.803], hereinafter referred to as the high ALOX15 group) showed significant improvement in 2-week rTNSS whether they received CM310 600 - 300 mg QW treatment or CM310 600 - 300 mg Q2W treatment compared with the control group. However, for the subjects with low expression of ALOX15 mRNA (ALOX15 mRNA < median [0.803], hereinafter referred to as the low ALOX15 group), after treatment with any dose of CM310 monoclonal antibody, there was no significant improvement in 2-week rTNSS compared with the control group. No such trend was observed for the other biomarkers. The changes in rTNSS scores of each subgroup of ALOX15 after 2 weeks of treatment with CM310 monoclonal antibody compared with the baseline are shown in Table 4.

[0085] Table 4 Changes in 2-week rTNSS scores compared with the baseline in the high ALOX15 group and the low ALOX15 group

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[0087] We further evaluated the improvement of the other clinical indicators in each subgroup of ALOX15 and found that the subjects in the high ALOX15 group who received CM310 treatment showed advantages in the improvement of nasal and ocular symptoms and quality of life. The changes in clinical indicators of each subgroup of ALOX15 after treatment with CM310 monoclonal antibody compared with the baseline are shown in Table 5.

[0088] Table 5 Changes in clinical indicators compared with the baseline in the high ALOX15 group and the low ALOX15 group

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[0104] Results Re-verification of Example 5

[0105] Patient Information and Study Design:

[0106] A total of 16 patients were included in this study. All patients needed to meet the following inclusion criteria: 1) aged 18 - 70 years; 2) received systemic corticosteroids (SCS) treatment within 2 years before the induction period, or had contraindications or intolerance to SCS treatment, or had undergone nasal polypectomy 6 months before the induction period; 3) nasal polyp score (NPS) was at least 5 points (at least 2 points in each nostril); 4) had moderate or severe nasal congestion (0 = asymptomatic, 1 = mild, 2 = moderate, 3 = severe), with an average weekly nasal congestion score (NCS) of 2 or 3 points, and any other symptoms such as anosmia or rhinorrhea; 5) patients needed to use nasal corticosteroids stably and continuously for at least 4 weeks before screening, and during the whole study, patients received MFNS as background treatment; 6) the percentage of peripheral blood eosinophils in all patients was ≥6.9% (without asthma) or ≥3.7% (with asthma); 2) through nasal polyp biopsy, the absolute eosinophil count in the tissue was ≥55 cells / high-power field or the eosinophil percentage was ≥27%.

[0107] Patients will be excluded if they have the following conditions: 1) have used an IL-4Rα antagonist within 10 weeks or 5 half-lives (whichever is longer) before randomization, or have used biologic therapy / systemic immunosuppressants within 8 weeks or 5 half-lives (whichever is longer), or have used anti-IgE monoclonal antibody within 130 days; 2) have undergone nasal polypectomy within 6 months before the screening / run-in period; 3) have received medium- and short-acting SCS treatment within 4 weeks before screening, or have received long-acting SCS treatment within 6 weeks; 4) have severe liver or kidney injury; 5) have received live or attenuated vaccines within 12 weeks before randomization, or plan to receive live and attenuated vaccines during the study; 6) patients known or suspected of immunosuppression, including but not limited to those with a history of invasive opportunistic infections (such as tuberculosis, histoplasmosis, listeriosis, coccidiosis, pulmonary cysts, aspergillosis), even if the infection has been resolved; 7) pregnant or lactating women, or women planning to become pregnant during the study; 8) any other medical or non-medical conditions that the researchers consider unsuitable for participation in the study. This study was approved by the Ethics Committee of Beijing Tongren Hospital, Capital Medical University. Written informed consent was obtained before patient participation.

[0108] After patients signed the informed consent form, they entered a 4-week screening / run-in period. During this period, all patients received MFNS nasal spray, 100 μg per nasal cavity per day (total daily dose was 200 μg); then patients received anti-IL-4Rα monoclonal antibody treatment for 16 weeks (the first subcutaneous injection was 600 mg, and then 300 mg every 2 weeks).

[0109] Sample detection:

[0110] Patients had nasal secretions and nasal polyps collected at baseline and 16 weeks of treatment for detecting the expression of ALOX15 and 15(S)-HETE in nasal secretions and nasal polyp tissues.

[0111] Obtaining nasal polyp tissues and detecting the expression level of ALOX15: Select typical lesion sites of nasal polyps, use a ethmoid sinus forceps or a punch forceps to obtain polyp tissues with a diameter of about 3 - 5 mm. Embed the patient's nasal polyp tissues in paraffin. Perform immunohistochemical staining on 4-μm tissue sections embedded in paraffin: Primary antibody: Anti-15 Lipoxygenase 1, ab244205, abcam; Secondary antibody: HRP-labeled. Examine under a microscope and collect and analyze images. After taking images with a bright-field optical microscope, perform immunohistochemical semi-quantitative analysis using ImageJ.

[0112] Obtaining and detecting 15(S)-HETE in nasal secretions: Place the trimmed and expanded sponge into the upper fornix of the left and right nasal cavities to collect nasal secretions and wait for 5 - 10 minutes. After removing the expanded sponge strips, place them in the corresponding left and right green 15 mL centrifuge tubes according to the left and right nasal cavities, add 0.5 mL of 0.9% normal saline, store at 4°C for 2 hours to obtain diluted nasal secretions, and centrifuge to obtain the supernatant of the secretions. Aliquot the supernatant and store it in an -80°C refrigerator, and uniformly perform 15(S)-HETE protein concentration detection later. Detection: Use an ELISA detection kit for 15(S)-HETE (#534721, Cayman).

[0113] Sample test results:

[0114] Before and after treatment with the anti-IL-4Rα monoclonal antibody CM310 in patients with ECRSwNP, immunohistochemical staining was used to evaluate the expression of ALOX15 protein, and ELISA was used to detect the expression of 15(S)-HETE protein( Figure 4 ).

[0115] The expressions of ALOX15 and 15(S)-HETE decreased after treatment with CM310 monoclonal antibody.

[0116] The above are the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. Use of a detection reagent for ALOX15 in the preparation of a kit for predicting or evaluating the efficacy of anti-IL-4Rα monoclonal antibody CM310 in the treatment of nasal mucosa type 2 inflammation-related nasal diseases.

2. The application according to claim 1, wherein The detection reagent for ALOX15 is a reagent for detecting the expression level of ALOX15.

3. The application according to claim 2, wherein The reagent for detecting the expression level of ALOX15 is a reagent for detecting the expression level of ALOX15 mRNA.

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