Traditional Chinese medicine preparation for treating cervical high-risk HPV persistent infection and application of traditional Chinese medicine preparation

Abstract

The invention relates to the technical field of virus infection treatment, in particular to a traditional Chinese medicine preparation for treating cervical high-risk HPV persistent infection and application of the traditional Chinese medicine preparation. The traditional Chinese medicine composition comprises the following components in parts by weight: 6-30 parts of radix aconiti carmichaeli (black prepared common monkshood daughter root) (decocted first), 3-15 parts of ginseng, 9-60 parts of polyporus umbellatus, 6-30 parts of fructus cnidii, 6-30 parts of fried semen brassicae, 6-30 parts of notopterygium root, 6-24 parts of radix bupleuri, 6-30 parts of fried green tangerine peel, 6-30 parts of motherwort and 6-30 parts of angelica sinensis. According to the constitution and syndrome differentiation of high-risk human papilloma virus (HR-HPV) persistent infection patients, a traditional Chinese medicine prescription is prepared on the basis of the traditional Chinese medicine yang-warming, cold-dispelling and dampness-eliminating theory, the yang-warming, cold-dispelling and dampness-eliminating treatment prescription is used for treating the persistent HPV infection and mild cervical lesion patients, and the traditional Chinese medicine composition can be used for treating the persistent HPV infection and mild cervical lesion patients on the basis of improving the internal environment of the body of the patient and boosting the healthy qi (autoimmunity). Therefore, evil (pathogenic microorganisms) are driven out.

Description

Technical Field

[0001] This invention relates to the field of viral infection treatment technology, specifically to a traditional Chinese medicine preparation for treating persistent infection with high-risk HPV of the cervix and its application. Background Technology

[0002] High-risk human papillomavirus (HR-HPV) infection is a major cause of cervical cancer, with 99.7% of cervical cancer patients also infected with HPV. Although some HPV infections can be cleared by the body's immune system, approximately 20%-30% persist, eventually developing into cervical intraepithelial neoplasia (CIN) and even cervical cancer. Studies show that if HR-HPV infection persists for more than a year, 21% of patients may develop high-grade cervical lesions (CIN II / III), also known as precancerous cervical lesions. Therefore, effective intervention for persistent high-risk HPV infection of the cervix is ​​crucial to reducing the incidence of precancerous cervical lesions and cervical cancer in women.

[0003] The WHO guidelines for screening and treatment of cervical precancerous lesions (2nd edition) recommend HPV DNA testing as the preferred method for cervical cancer screening. If HPV DNA testing is positive, cytological or colposcopy-guided histological examination should be performed. If the colposcopy-guided histological examination is negative or shows low-grade cervical intraepithelial neoplasia (CIN I), HPV testing should be repeated in 6-12 months. If the colposcopy-guided histological examination shows high-grade cervical intraepithelial neoplasia (CIN II / III) or carcinoma in situ, ablation or surgical treatment should be performed. It can be seen that for HPV infection and low-grade cervical lesions, clinical observation is the primary approach, and there are no effective treatments. Once the condition progresses to high-grade cervical lesions or cervical cancer, surgical treatment is necessary.

[0004] In summary, there is currently no effective treatment for HPV infection. Treatment primarily relies on boosting immunity for self-clearance, and some vaginal topical medications are also available. However, after HPV infection, it can enter cervical cells and even integrate with cervical cell DNA, thus topical medications are not very effective. Chinese patent CN119033874A discloses a traditional Chinese medicine composition and preparation method for treating HPV infection and cervical precancerous lesions. This method includes both oral and topical formulations. The oral formulation is as follows: Angelica sinensis 6-12g, Paeonia lactiflora 6-15g, Rehmannia glutinosa 10-15g, Ligusticum chuanxiong 3-10g, Astragalus membranaceus 10-30g, Codonopsis pilosula 10-30g, Panax ginseng 3-9g. The formula contains 15-30 grams of Dioscorea opposita, 6-15 grams of Forsythia suspensa, 3-12 grams of Phellodendron chinense, 5-10 grams of Sophora flavescens, 10-15 grams of Dictamnus dasycarpus, 6-15 grams of Lonicera japonica, and 10-15 grams of Taraxacum mongolicum. It mainly functions to clear heat and eliminate dampness and toxins, tonify the spleen and replenish qi, nourish blood and promote blood circulation, and nourish yin and replenish essence. However, the patent does not provide verification data, and in clinical practice, we have found that most patients with persistent HPV infection have the pathogenesis characteristics of "insufficient yang qi and cold-dampness obstruction", which is difficult to meet the needs of clinical treatment. Summary of the Invention

[0005] The technical problem this invention aims to solve is that there are currently no effective treatments for HPV infection. Treatment mainly relies on boosting the immune system for self-clearance, and some topical vaginal medications are also available. However, after HPV infection, it can enter cervical cells and even integrate with cervical cell DNA, thus making topical medications less than ideal.

[0006] To address the aforementioned problems, this invention provides a traditional Chinese medicine preparation for treating persistent high-risk HPV infection of the cervix and its application. Based on the constitution and syndrome differentiation of patients with persistent high-risk human papillomavirus (HR-HPV) infection, and based on the TCM principles of warming yang, dispelling cold, and eliminating dampness, a TCM formula is formulated. This formula is used to treat patients with persistent HR-HPV infection and mild cervical lesions by improving the patient's internal environment and boosting their own vital energy (immunity), thereby expelling pathogenic microorganisms.

[0007] To achieve the above objectives, the technical solution of this invention is: a traditional Chinese medicine preparation for treating persistent infection of high-risk HPV in the cervix, wherein the components of the traditional Chinese medicine composition by weight are as follows: Aconitum carmichaelii (black aconite slices) (decocted first) 6-30 parts, ginseng 3-15 parts, Polyporus umbellatus 9-60 parts, Cnidium monnieri 6-30 parts, stir-fried white mustard seed 6-30 parts, Notopterygium incisum 6-30 parts, Bupleurum chinense 6-24 parts, stir-fried Citrus reticulata peel 6-30 parts, Leonurus japonicus 6-30 parts, and Angelica sinensis 6-30 parts.

[0008] Further, the preferred ingredients are: 9 parts of Aconitum carmichaelii (black slices) (decocted first), 6 parts of ginseng, 9 parts of Polyporus umbellatus, 9 parts of Cnidium monnieri, 9 parts of Sinapis alba, 9 parts of Notopterygium incisum, 9 parts of Bupleurum chinense, 9 parts of stir-fried Citrus reticulata peel, 15 parts of Leonurus japonicus, and 9 parts of Angelica sinensis.

[0009] Traditional Chinese medicine (TCM) does not have a specific disease name for "HPV infection," but it considers it to be an external invasion of damp-heat toxins. On one hand, patients are prone to external damp-heat toxins due to long-term weakness of their vital energy; on the other hand, the long-term invasion of damp-heat toxins within the body further damages the body's vital energy, leading to the disease. Therefore, "deficiency of vital energy and accumulation of toxins" is the basic pathogenesis of this disease. Clinical manifestations often include: aversion to cold, cold lower back and abdomen, fatigue, heaviness in the limbs, chronic insomnia or excessive sleepiness, loss of appetite, lower abdominal distension, loose or sticky stools, vaginal itching, copious and cloudy leukorrhea, irregular menstruation or dysmenorrhea. These patients often present with a pale tongue with teeth marks, a white and greasy coating at the root of the tongue, dark sublingual veins, and dark lips. The pulse is characterized by a deep, wiry, and thready pulse, with a deep pulse at the cun position.

[0010] Upon examination and investigation, the etiology of this disease is found to be both internal injury from overwork and external invasion of pathogenic factors. The location of the disease is in the cervix, and its nature is characterized by deficiency of the root and excess of the branch. The combined factors of etiology, location, and nature of the disease indicate that the pathogenesis of this type of disease is internal yang deficiency and obstruction by cold and dampness. The prescribed formula is a warming yang, dispersing cold, and eliminating dampness formula, consisting of 9 grams each of Aconitum carmichaelii (black aconite slices) (decocted first), Polyporus umbellatus, Cnidium monnieri, stir-fried white mustard seed, Notopterygium incisum, Bupleurum chinense, stir-fried Citrus reticulata peel, and Angelica sinensis, 6 grams of ginseng, and 15 grams of Leonurus japonicus. The principal herbs are Aconitum carmichaelii (black aconite slices) and ginseng, which together warm and tonify the primordial yang to support the body's resistance and expel pathogenic factors. The assistant herbs are Polyporus umbellatus, Cnidium monnieri, stir-fried white mustard seed, and Notopterygium incisum, which expel cold, dampness, phlegm, and parasites, assisting the principal herbs in achieving the effects of warming yang, dispersing cold, and promoting diuresis. The adjuvant herbs are Bupleurum chinense and stir-fried Citrus reticulata peel, which soothe stagnation, regulate qi, and unblock the meridians, enhancing the efficacy of the principal and assistant herbs. The guiding herbs are Leonurus japonicus and Angelica sinensis, which carry the other herbs directly to the affected area.

[0011] Modern pharmacology of the formula for warming yang, dispelling cold, and eliminating dampness: Aconite is primarily used to treat wind-cold, dispel cold and dampness, and eliminate deep-seated cold in the internal organs, while also resolving stagnation and promoting circulation. Modern pharmacological research suggests that aconite possesses anti-inflammatory, analgesic, immune-enhancing, and anti-tumor pharmacological effects. Its anti-cancer mechanism is mainly related to enhancing the body's cellular immune function, inducing tumor cell apoptosis, and regulating oncogene expression.

[0012] Ginseng is a powerful tonic for vital energy, and it strengthens the spleen and lungs. Studies have found that ginsenosides, an extract of ginseng, can inhibit HPV, promote apoptosis in cervical cancer cells, and regulate their autophagy.

[0013] Cnidium monnieri is often used to treat gynecological diseases. Experiments have shown that Cnidium monnieri has the effect of antagonizing osteoporosis caused by hormones, improving the body's immunity, regulating the levels of SOD and sex hormones (E2 / T), and has anti-allergic and antipruritic effects. Osthol also has strong anti-cancer activity against cervical cancer.

[0014] Notopterygium root is mainly used to treat wind-cold-dampness arthralgia and to dispel wind-evil between the kidneys. Notopterygium root extract has a certain killing effect on pathogenic microorganisms, and Notopterygium root volatile oil has a significant effect on improving fever caused by endotoxins.

[0015] Bupleurum soothes the liver and relieves depression, raises yang energy, and has a targeted effect, guiding the medicine to the liver meridian (which circulates around the genitals).

[0016] Stir-fried white mustard seeds can eliminate phlegm caused by coldness and dampness in the skin and membranes. Pharmacological studies have reported that its aqueous solution has an inhibitory effect on skin fungi in vitro.

[0017] Green tangerine peel is a widely used, safe and effective drug in clinical practice. Clinical observations have confirmed that it has significant therapeutic effects on reproductive system diseases such as chronic pelvic inflammatory disease.

[0018] Polyporus umbellatus is mainly used to treat malaria, detoxify parasites, and promote urination. Modern Chinese Materia Medica suggests that Polyporus umbellatus can detoxify cancer. Studies have shown that Polyporus umbellatus polysaccharides promote IL-2 synergistic induction of human peripheral monocytes into cells with high tumor-killing activity. Polyporus umbellatus has immunomodulatory and anti-tumor effects.

[0019] Motherwort invigorates blood circulation, promotes menstruation, breaks up blood stasis, clears the meridians, expels stillborn fetuses, and destroys the placenta... It is a good medicine for women. Modern pharmacological research has confirmed that its extract, leonurine, has a protective effect on the reproductive system.

[0020] Angelica sinensis nourishes blood and liver... it invigorates the subtle meridians and relieves coldness in the limbs... it is most effective in warming the meridians. Modern research has found that its pharmacological activities are mainly concentrated in anti-inflammatory, analgesic, antiviral, antitumor, and anti-allergic aspects.

[0021] In summary, persistent infection with high-risk HPV is a major risk factor for cervical cancer, significantly impacting women's physical and mental health. Currently, modern medicine lacks particularly effective treatments and primarily employs lifestyle interventions, including guiding patients to maintain regular sleep patterns, balanced nutrition, and appropriate exercise, hoping to improve immunity through lifestyle changes (as this is a common approach, using a control group is reasonable). This study, based on traditional Chinese medicine (TCM) diagnosis of patients with persistent high-risk HPV infection, formulated a TCM prescription and conducted a randomized controlled trial to evaluate the effectiveness of TCM treatment in eliminating HPV infection and the seroconversion rate of various high-risk HPV types compared to conventional lifestyle interventions. This aims to explore effective treatment methods for persistent high-risk HPV infection.

[0022] Furthermore, the traditional Chinese medicine preparation also includes pharmaceutically acceptable excipients.

[0023] Furthermore, the dosage forms of the traditional Chinese medicine preparations include decoctions, granules, pills, capsules, tablets, powders, or oral liquids.

[0024] The application of the traditional Chinese medicine preparation in the preparation of drugs for treating persistent infection with high-risk HPV of the cervix.

[0025] The preparation process of granules is as follows: Weigh the prescribed amount of Chinese medicinal materials, clean and pulverize them into coarse powder, and pass them through a 10-mesh sieve. Soak the powder in 10 times its volume of purified water for 30 minutes, then reflux for 1.5 hours and filter. Add 8 times its volume of purified water to the residue and reflux for 1.0 hour, then filter again. Combine the two filtrates. After centrifugation to remove impurities, concentrate the filtrate at 65℃ and a vacuum of -0.085 MPa to a relative density of 1.22 (60℃). Spray dry the filtrate at an inlet temperature of 175℃ and an outlet temperature of 90℃ to obtain a dry powder, which is then passed through an 80-mesh sieve. Add 10%–20% dextrin and 5%–10% lactose according to the amount of dry powder, mix thoroughly, and prepare a soft mass using 50% ethanol as a wetting agent. Granulate the granules through a 14-mesh sieve. Boil and dry the wet granules at 62℃ until the moisture content is 4.2%, then granulate using a 14-mesh sieve. After granulation, the granules are placed in a three-dimensional mixer and mixed for 25 minutes. They are then packaged into 5 g bags, sealed, and the self-formulated Chinese medicine granules are obtained.

[0026] The beneficial effects of this invention are as follows: This traditional Chinese medicine formula for warming yang, dispelling cold, and eliminating dampness targets persistent high-risk HPV infection in cervical cervical stenosis caused by yang deficiency and cold-dampness. Compared to lifestyle interventions alone, it significantly improves the HPV clearance efficiency and negative conversion rate, improves cervical cytological abnormalities, and has a clearing effect on multiple HPV subtypes, especially achieving complete clearance of types 39, 58, and 66. The formula is scientifically formulated and has been verified by modern pharmacology to have immunomodulatory and antiviral effects. It is safe to use with no serious adverse events, providing a safe and effective non-invasive treatment for persistent HPV infection, which can prevent the progression of the disease to cervical cancer and reduce the incidence of cervical cancer. Attached Figure Description

[0027] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0028] Figure 1 Comparison of HPV clearance efficacy between the two groups.

[0029] Figure 2 Secondary outcome: TCT negative conversion rate.

[0030] Figure 3 Secondary outcome: Comparison of clearance rates for different HPV subtypes.

[0031] Figure 4 Forest plot of subgroup analysis. Detailed Implementation

[0032] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0033] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or intermediate value within a stated range, and any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0034] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0035] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be readily apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0036] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0037] Unless otherwise specified, the experimental methods used in the following examples are conventional methods; the materials and reagents used are commercially available or can be prepared by existing methods.

[0038] The following examples all use the following traditional Chinese medicine preparations: A traditional Chinese medicine preparation for treating persistent infection with high-risk HPV of the cervix, comprising, by weight, the components of the traditional Chinese medicine composition. The following is the formula: 9 parts of Aconitum carmichaelii (black slices) (decocted first), 6 parts of ginseng, 9 parts of Polyporus umbellatus, 9 parts of Cnidium monnieri, 9 parts of stir-fried white mustard seed, 9 parts of Notopterygium incisum, 9 parts of Bupleurum chinense, 9 parts of stir-fried Citrus reticulata peel, 15 parts of Leonurus japonicus, and 9 parts of Angelica sinensis.

[0039] Example 1: Preliminary Experimental Study (1) Research subjects Fifteen patients with persistent high-risk HPV infection of the cervix were selected and treated with traditional Chinese medicine; another 20 patients with the same type of high-risk HPV infection were selected as controls and only received lifestyle guidance.

[0040] (2) Intervention methods The experimental group was given the oral administration of the formula of the present invention, which consists of 9 grams each of Aconitum carmichaelii (black aconite slices) (decocted first), Polyporus umbellatus, Cnidium monnieri, stir-fried white mustard seed, Notopterygium incisum, Bupleurum chinense, stir-fried Citrus reticulata peel, and Angelica sinensis, 6 grams of ginseng, and 15 grams of Leonurus japonicus, and was taken according to the prescribed method; the control group was only given lifestyle guidance on regular work and rest, balanced nutrition, and appropriate exercise.

[0041] (3) Research Results At the 6-month follow-up after treatment, among the 15 patients in the experimental group, 5 had all their original high-risk HPV subtypes turned negative (cured), 6 had partially turned negative (significantly effective), and 4 did not turn negative (ineffective). Effectiveness includes both cure and significant effect. Based on this, the total effective rate was 73.3%. In the control group, after 6 months of follow-up, the total effective rate was only 36%. The efficacy of the experimental group was significantly better than that of the control group.

[0042] Example 2: Randomized Controlled Clinical Trial I. Experimental Design (1) Research Design and Research Subjects This prospective, randomized, open-label, parallel-controlled clinical trial was conducted from May to November 2024 at the Department of Traditional Chinese Medicine, Beijing Anzhen Hospital, Capital Medical University, to evaluate the efficacy and safety of a warming-yang-dispersing-cold-removing-dampness formula combined with lifestyle intervention alone in women with persistent high-risk HPV infection. The study protocol was approved by the institution's ethics committee, and all participants signed informed consent forms.

[0043] For participant selection, the inclusion criteria were: women aged 24-77 years with persistent HR-HPV infection for ≥12 months. All participants were diagnosed by a Traditional Chinese Medicine (TCM) practitioner, and only patients with Yang deficiency and cold-dampness type were included (main characteristics include aversion to cold, cold lower abdomen, fatigue, heaviness in the body, shortness of breath, reluctance to speak, chronic insomnia or somnolence, poor appetite, edema of the limbs, lower abdominal distension, loose or sticky stools, irregular menstruation or dysmenorrhea, pale tongue with teeth marks, white and greasy coating at the root of the tongue, and deep, wiry, and thready pulse). Eligible participants had persistent infection with one or more of the 14 HR-HPV subtypes (HPV 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, or 68), confirmed by two consecutive positive HPV DNA tests at least ≥12 months apart. CIN I patients who were untreated were also eligible for inclusion.

[0044] Exclusion criteria: Pregnancy or lactation; history of cervical cancer treatment; serious cardiovascular, pulmonary, hepatic, renal, hematologic diseases, or malignant tumors; immune system diseases or immunosuppressive therapy; use of corticosteroids, antiviral drugs, or drugs acting on the immune system within the past 3 months; having undergone loop electrosurgical excision procedure (LEEP) or cold knife conization (CKC); currently taking traditional Chinese medicine for other diseases; having vaginal infections requiring treatment; allergy to the prescribed traditional Chinese medicine; having other uncontrolled diseases that the investigator deems unsuitable for participation; any situation that the investigator believes may increase the risk to the subject or interfere with the trial results.

[0045] Withdrawal criteria: Subject requests to withdraw from clinical trial; serious violation of study protocol; subject becomes pregnant; serious adverse reaction occurs and the investigator determines that the subject needs to withdraw from clinical trial; there is evidence of disease progression in the subject.

[0046] (2) Randomization and intervention measures Subsequently, 126 participants were randomly assigned 1:1 to either the treatment group (n=63) or the control group (n=63) using computer-generated random numbers. Randomization was performed by an independent statistician, using sequential numbering, opacity, and sealed envelopes for allocation concealment. Participants and researchers were not blinded; however, the assessors of HPV DNA testing and cytology results were blinded regarding the group assignments. Specifically, participants in the treatment group took one dose of the concentrated granules of the warming and dispersing cold-dispelling formula of this invention, twice daily with warm water half an hour after meals, starting 2–3 days after menstruation ended, for 20 days each month, for 3 consecutive months, discontinuing medication during menstruation. The control group received only lifestyle interventions. Both groups received the same lifestyle guidance, including regular sleep patterns, balanced nutrition, appropriate exercise, use of condoms during sexual intercourse to avoid cross-infection, and smoking cessation if applicable. All participants avoided the use of antiviral drugs, immunomodulators, and vaginal preparations during the study period.

[0047] (3) Observation indicators The primary endpoint of this study was the overall response rate to HPV elimination at 6 months post-treatment, defined as complete seroconversion (all baseline HR-HPV subtypes becoming negative) or significant response (reduction in detected subtypes). Ineffectiveness was defined as the persistence of all baseline subtypes. Differences between groups were quantified using odds ratios (OR) and 95% confidence intervals (CI). In addition, secondary endpoints included: (1) subtype-specific HPV seroconversion rates for each genotype (HPV types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68); and (2) cytological seroconversion rates in subjects with baseline TCT abnormalities who achieved NILM status. The pre-defined subgroup analyses included: age (<40 years vs ≥40 years), TCT status (NILM vs abnormal), pathology (normal / no abnormality vs CIN positive), HPV 16 / 18 infection status (including 16 / 18 types vs non-16 / 18 types), and infection pattern (single subtype vs multiple infections).

[0048] (4) Clinical assessment In terms of clinical assessment, baseline evaluation included demographic data (age, weight, ethnicity), gynecological examination, HR-HPV DNA genotyping, liquid-based thin-layer cytology (TCT), colposcopy, and cervical biopsy when necessary, as well as laboratory tests (complete blood count, liver function, kidney function). HPV genotyping used a validated PCR-based kit capable of detecting 14 HR-HPV subtypes. Cytological specimens were analyzed using the ThinPrep liquid-based cytology method, interpreted according to the 2014 Bethesda classification system. Colposcopy followed a standard protocol, with biopsies taken from suspicious areas, or from the 3, 6, 9, and 12 o'clock positions when no lesions were found. Histological specimens were evaluated by at least two pathologists, with disagreements resolved through discussion. During follow-up, at the 6-month follow-up, evaluation included gynecological examination, repeated HPV DNA genotyping, TCT, colposcopy with cervical biopsy when necessary, vital signs, and laboratory tests. Adverse events were systematically recorded and graded according to CTCAE version 5.0 during the study.

[0049] (5) Statistical analysis methods For statistical analysis, sample size was calculated based on expected efficacy rates of 65% (treatment group) and 35% (control group), with α=0.05 and a power of 80%. Considering a 15% dropout rate, the target enrollment was 126 subjects. The full analysis set (including all randomly assigned subjects with at least one post-baseline efficacy evaluation) was used as the primary population for analysis, following the intention-to-treat principle. Continuous variables were analyzed using t-tests or Mann-Whitney U tests; categorical variables were analyzed using chi-square tests or Fisher's exact test. The primary endpoint was compared between groups using chi-square tests, and treatment efficacy was quantified using logistic regression models to calculate OR and 95% CI. Subgroup analyses calculated efficacy rates and 95% CIs within pre-defined strata, and forest plots were used to visualize the consistency of treatment efficacy. All analyses were performed using R version 4.1.1 software. A two-sided P < 0.05 was considered statistically significant.

[0050] II. Results (1) Study subjects and baseline characteristics A total of 126 patients with persistent high-risk HPV infection were enrolled and randomly assigned to receive either a warming and dispersing formula (n=63) or a lifestyle intervention alone (n=63). During the 6-month follow-up period, 9 patients in the treatment group and 11 patients in the control group withdrew. Therefore, 54 patients in the treatment group and 52 patients in the control group completed the study and were included in the efficacy analysis. The baseline characteristics of the two groups were well balanced (Table 1). The mean age of the treatment group (39.9±12.5 years) was comparable to that of the control group (39.7±14.1 years). Similarly, no significant differences were observed in baseline TCT results (NILM: 51.9% vs 50.0%; abnormal: 48.1% vs 50.0%), pathological results (normal / no abnormalities: 31.5% vs 32.7%; CIN I: 68.5% vs 67.3%), HPV 16 / 18 infection status (including 16 / 18: 20.4% vs 21.2%; non-16 / 18: 79.6% vs 78.8%), or infection type (single infection: 68.5% vs 73.1%; multiple infections: 31.5% vs 26.9%).

[0051] Table 1. Comparison of baseline characteristics between the treatment group and the control group

[0052] (2) Primary therapeutic endpoint Regarding the primary outcome, the HPV clearance rate in the treatment group was significantly better than that in the control group (Table 2). Figure 1In the treatment group, 66.7% (36 / 54) of patients achieved complete HPV clearance (negative clearance), compared to 32.7% (17 / 52) in the control group. Partial clearance (significant effect) was observed in 11.1% (6 / 54) of patients in the treatment group and 9.6% (5 / 52) in the control group. The overall response rate was defined as complete clearance plus significant effect, and the treatment group (77.8%, 42 / 54) was significantly higher than the control group (42.3%, 22 / 52), with an odds ratio (OR) of 4.77 (95% CI: 2.05–11.11, P = 0.0004).

[0053] Table 2. Main outcomes: Comparison of HPV clearance effectiveness (P<0.01)

[0054] (3) Secondary therapeutic endpoints Furthermore, the analysis of secondary key indicators supports the therapeutic effect of the warming-yang-dispersing-cold-removing-dampness formula. Figure 2 In patients with abnormal baseline TCT, the TCT seroconversion rate in the treatment group (42.3%, 11 / 26) was significantly higher than that in the control group (11.5%, 3 / 26, P=0.0266).

[0055] Further analysis showed that subtype-specific HPV seroconversion rates were analyzed for all 11 high-risk HPV genotypes present in the two studies. Figure 3 The treatment group showed consistently high clearance rates across most HR-HPV subtypes. Complete clearance (100%) was achieved for HPV39 (6 / 6 vs 5 / 12, 42%), HPV58 (4 / 4 vs 4 / 9, 44%), and HPV66 (2 / 2 vs 0 / 2, 0%). High clearance rates were also observed for HPV59 (75% vs 0%), HPV56 (71% vs 22%), HPV31 (67% vs 0%), and HPV52 (67% vs 28%). Notably, HPV52 was the most common subtype, with 21 cases in the treatment group and 18 in the control group, representing a 39 percentage point difference in clearance rate. For clinically significant high-risk types, the seroconversion rate for HPV16 was 56% (5 / 9) compared to 25% (3 / 12), while for HPV18 it was 50% (1 / 2) compared to 0% (0 / 2). HPV45 did not show seroconversion in either group (0 / 2 and 0 / 4, respectively).

[0056] (4) Subgroup analysis To assess the consistency of treatment outcomes, pre-specified subgroup analyses showed that treatment outcomes were consistent across all patient subgroups examined. Figure 4Regardless of age (<40 years: 28 / 36 vs 10 / 30, OR=7.00, 95% CI: 2.35-20.87; ≥40 years: 14 / 18 vs 12 / 22, OR=2.92, 95% CI: 0.72-11.74), TCT status (NILM: 21 / 28 vs 11 / 26, OR=4.09, 95% CI: 1.29-13.00; abnormal: 21 / 26 vs 11 / 26, OR=5.73, 95% CI: 1.64-19.94), pathology (normal / no abnormality: 13 / 17 vs 8 / 17, OR=3.66, 95% CI: 0.84-15.91; CIN positive: 29 / 37 vs 14 / 35, OR=5.44, 95%), the incidence rate was significantly lower. Treatment benefits were observed regardless of HPV 16 / 18 status (including 16 / 18: 6 / 11 vs 2 / 11, OR=5.40, 95% CI: 0.78-37.51; non-16 / 18: 36 / 43 vs 20 / 41, OR=5.40, 95% CI: 1.96-14.90) or infection type (single infection: 29 / 37 vs 14 / 38, OR=6.21, 95% CI: 2.23-17.29; multiple infections: 13 / 17 vs 8 / 14, OR=2.44, 95% CI: 0.52-11.39). All subgroup point estimates supported the treatment group, with odds ratios consistently exceeding 1. The forest plot showed that the overall treatment effect (OR=4.77, 95% CI: 2.05-11.11) remained robust across different patient characteristics, suggesting that the treatment intervention has broad applicability.

[0057] (5) Safety and tolerability Regarding safety, the warming and dispersing cold and dampness-removing formula demonstrated good safety throughout the study period (Table 3). All 54 patients in the treatment group and all 52 patients in the control group completed the full course of treatment. No adverse events were reported in the treatment group (0 / 54, 0%), while one adverse event occurred in the control group (1 / 52, 1.9%). The difference in the incidence of adverse events was not statistically significant (P=0.491). Importantly, no serious adverse events or treatment discontinuation due to adverse events were observed in either group, indicating that the traditional Chinese medicine intervention was well-tolerated.

[0058] Table 3 Security Analysis

[0059] III. Conclusion: Our primary findings demonstrate that the warming and dispersing formula of this invention is significantly superior to lifestyle intervention alone in HPV clearance efficacy, with good safety and tolerability. The seroconversion rate observed in the control group was consistent with the natural clearance rate in observational studies. However, the treatment group showed a significantly higher seroconversion rate above this baseline, providing evidence of therapeutic benefit attributable to traditional Chinese medicine intervention. These findings are consistent with recent evidence that drug intervention can effectively enhance viral clearance beyond the natural course of the disease. Subgroup analysis of different patient subgroups showed consistent treatment benefit across all patient characteristics examined. The treatment response pattern between single and multiple HPV infections is consistent with existing evidence that multiple co-infections present unique clinical challenges. Subtype-specific analysis revealed heterogeneity in seroconversion patterns among different HPV genotypes. The relatively low seroconversion rate observed for HPV16, the most carcinogenic subtype globally, may reflect characteristics of viral genome integration.

[0060] From a mechanistic perspective, the observed therapeutic effects can be attributed to the immunomodulatory and antiviral properties of the formula's components, consistent with the principles of Traditional Chinese Medicine (TCM) in restoring the body's defensive immune capacity. Modern pharmacological studies have demonstrated that ginsenosides exhibit anticancer activity and immunomodulatory effects against cervical cancer cells, Cnidium monnieri and its component osthol exhibit immune-enhancing and apoptosis-inducing properties, and Aconitum carmichaelii-based drugs exhibit anti-inflammatory and immunomodulatory effects. These pharmacological properties provide a reasonable biological basis for the observed clinical benefits. Furthermore, secondary endpoints support the treatment benefit: the treatment group showed significant improvement in both baseline cytological abnormalities and CIN patients, which is significant for preventing disease progression. CIN represents a critical stage in the progression from persistent HPV infection to invasive cancer, and pathological improvement suggests benefit from the regression of existing lesions.

[0061] In summary, this randomized controlled trial demonstrated that the warming and dispersing cold-dampness formula was significantly superior to lifestyle intervention alone, suggesting that rigorously evaluated traditional Chinese medicine methods can expand treatment options for persistent HR-HPV infection and fill the key gap between screening detection and surgical treatment. However, larger sample sizes and longer follow-up studies are needed to establish the durability of efficacy and its clinical impact on cervical cancer prevention.

[0062] The above description is merely a preferred embodiment of the present invention and does not constitute any limitation on the present invention. Any simple modifications, alterations, and equivalent transformations made to the above embodiments based on the technical essence of the present invention shall still fall within the protection scope of the present invention.

Claims

1. A traditional Chinese medicine preparation for treating persistent infection with high-risk HPV of the cervix, characterized in that, The components of the traditional Chinese medicine composition by weight are as follows: 6-30 parts of black aconite slices (decocted first), 3-15 parts of ginseng, 9-60 parts of poria cocos, 6-30 parts of cnidium monnieri, 6-30 parts of stir-fried white mustard seed, 6-30 parts of notopterygium incisum, 6-24 parts of bupleurum chinense, 6-30 parts of stir-fried green tangerine peel, 6-30 parts of motherwort, and 6-30 parts of angelica sinensis.

2. The traditional Chinese medicine preparation as described in claim 1, characterized in that, The components of the traditional Chinese medicine composition by weight are as follows: 9 parts of black aconite root slices (decocted first), 6 parts of ginseng, 9 parts of poria cocos, 9 parts of cnidium monnieri, 9 parts of white mustard seed, 9 parts of notopterygium incisum, 9 parts of bupleurum chinense, 9 parts of stir-fried green tangerine peel, 15 parts of motherwort, and 9 parts of angelica sinensis.

3. The traditional Chinese medicine preparation as described in claim 1, characterized in that: The traditional Chinese medicine preparation also includes pharmaceutically acceptable excipients.

4. The traditional Chinese medicine preparation as described in claim 1, characterized in that: The dosage forms of the traditional Chinese medicine preparations include decoctions, granules, pills, capsules, tablets, powders, or oral liquids.

5. The use of the traditional Chinese medicine preparation according to any one of claims 1-4 in the preparation of a drug for treating persistent infection of high-risk HPV in the cervix.

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