Plant compound gynecological anti-HPV gel preparation based on multi-target intervention

Abstract

The invention discloses a multi-target intervention-based plant compound gynecological anti-HPV (human papillomavirus) gel preparation, which comprises two parts, namely a compound plant extracting solution and an aqueous gel carrier, the compound extracting solution is derived from rheum officinale, scutellaria baicalensis, coptis chinensis, golden cypress, radix sophorae flavescentis, fructus cnidii, alum, gallnut, common carpesium fruit, radix angelicae, cortex dictamni, fructus kochiae, frankincense and myrrh; the anti-HPV gel preparation is obtained by mixing and stirring according to the proportion. Aiming at an intervention window that a patient is continuously infected but does not deteriorate to an operation stage, intervention is carried out by adopting an action mechanism of local epithelial micro-ecological restoration and mucosal immunity, E6 / E7 oncogenic protein expression is inhibited, L1 capsid assembly is damaged, key nodes of a virus life cycle are covered, and escape risks are reduced by multi-target collaborative anti-HPV (Human Papilloma Virus); by inhibiting an NF-kB / EMT signal axis, the cervical lesion progress risk is reduced; the safety is high and the curative effect is obvious.

Description

Technical Field

[0001] The present invention belongs to the technical field of gel preparations, and in particular relates to a plant compound anti-HPV gel preparation for gynecological use based on multi-target intervention. Background Art

[0002] Human papillomavirus (HPV) is a group of non-enveloped, double-stranded DNA viruses belonging to the family Papillomaviridae. To date, over 200 HPV genotypes have been identified. Based on their carcinogenic risk, they are divided into low-risk and high-risk types. Low-risk HPV types (such as HPV-6 and HPV-11) primarily cause benign lesions such as genital warts, while high-risk HPV types (such as HPV-16, HPV-18, HPV-31, and HPV-33) are closely associated with the development of various malignancies, particularly cervical cancer. Types 16 and 18 cause over 70% of cervical cancer cases worldwide. The World Health Organization (WHO) classifies the following genotypes as high-risk oncogenic: genotypes 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68 [McBride, AA (2022) Human papillomaviruses: diversity, infection and host interactions. Nat. Rev. Microbiol. 20, 95–108] [Gheit, T. (2019) Mucosal and cutaneous human papillomavirus infections and cancer biology. Front. Oncol. 9, 355]. Genotypes 16 and 18 cause over 70% of cervical cancer cases worldwide. The HPV genome consists of three functional regions: the LCR (upstream regulatory region) is a non-coding region that plays a key role in viral DNA replication and transcriptional regulation. The other two regions are the "early region" and the "late region", which together encode eight open reading frames (ORFs), which are viral functional proteins.

[0003] Most sexually active people will be infected with HPV at some point in their lives. While most HPV infections are transient and cleared by the body's immune system within a few months, persistent infection with some high-risk HPV types can lead to precancerous lesions and, subsequently, malignant tumors. HPV infection is closely linked to a variety of diseases, including cervical cancer: nearly all cases of cervical cancer are linked to high-risk HPV infection. HPV-16 and HPV-18 account for approximately 70% of cervical cancer cases; other genital cancers: HPV infection is also associated with genital cancers such as vulvar cancer, vaginal cancer, penile cancer, and anal cancer; head and neck cancers: high-risk HPV infection is also closely linked to the development of head and neck squamous cell carcinomas, such as oropharyngeal cancer; and genital warts: low-risk HPV infection can cause genital warts (condyloma acuminata), impacting patients' quality of life.

[0004] Currently, the primary means of preventing HPV infection are vaccination and regular screening. HPV vaccines have been proven safe and effective in preventing HPV infection and related cancers. However, in China, HPV vaccine rollout has been relatively slow, leaving many women unvaccinated. Furthermore, HPV vaccines are limited to preventive measures and have no therapeutic effect on those already infected. For women already infected with HPV, interferon gels are commonly used, but these can have significant side effects and poor compliance. Conservative treatments, such as traditional Chinese medicine (TCM), are often used, but the mechanisms of action of most TCM gels remain unclear, leaving their composition and targets unclear. This means there is a lack of antiviral drugs that act directly on the cervical mucosa and target multiple targets. Clinically, a "watch and wait" approach is often adopted, with intervention only initiated after precancerous lesions develop, typically through ablation or surgery. These procedures can increase the risk of future complications such as difficulty conceiving and miscarriage. Therefore, effective antiviral treatments are currently lacking, necessitating the development of effective treatments for HPV infection.

[0005] More specifically, in some infected women, the HPV virus persists, and approximately 10% of those infected with HPV develop signs of cervical precancerous lesions or cancer. Persistent infection is not the same as ordinary infection; it is the true risk factor. We target the intervention window of persistent infection, but before the disease progresses to the point of requiring surgery, by employing a combination of local epithelial microecological restoration and mucosal immunity.

[0006] The present invention plans to develop a traditional Chinese medicine compound gel derived from plant components based on modern pharmacological validation, targeting E6 / E7 gene expression, viral DNA integration, immune escape and local microecological imbalance, and providing an auxiliary treatment plan suitable for people after HPV infection, that is, those in the observation period of cervical lesions (CIN I~II) or those who are HPV-positive but without lesions. Summary of the Invention

[0007] In view of the shortcomings of the existing technology, the present invention provides a plant compound anti-HPV gel preparation for gynecology based on multi-target intervention.

[0008] To achieve the above object, the present invention adopts the following technical solutions:

[0009] A plant compound anti-HPV gel preparation for gynecological use based on multi-target intervention, consisting of a compound plant extract and an aqueous gel carrier.

[0010] The compound plant extract is derived from: rhubarb, scutellaria, coptis root, phellodendron, sophora flavescens, cnidium monnieri, alum, gallnut, scutellaria baicalensis, angelica dahurica, dandelion bark, kochia scoparia, frankincense and myrrh;

[0011] The aqueous gel carrier consists of glycerol, purified water, propylene glycol, carbomer and alginate.

[0012] Preferably, the ratio of the components of the compound plant extract is: m Cnidium monnieri: m Alum: m Dictamnus cortex: m Kochia scoparia: m Frankincense: m Myrrh: m Gallnut = 1:1:1:1:1:1:2; m Gastrodia elata: m Angelica dahurica = 1:1; m Rhubarb: m Scutellaria baicalensis: m Coptis chinensis: m Phellodendron amurense: m Sophora flavescens = 1:1:1:1:1:1.

[0013] Furthermore, m Dictamnus cortex: m Galla chinensis: m Angelica dahurica: m Scutellaria baicalensis = 1:2:1.5:5.0.

[0014] By adopting the above technical scheme, the efficacy and function of each component of the plant extract, such as Emodin: inhibits E6 / p53 binding and blocks viral integration; Baicalin: downregulates E7 expression and interferes with the AP-1 pathway; Matrine: interferes with L1 capsid protein self-assembly; Ellagic acid: affects viral DNA methylation and enhances p21 expression; Osthole: activates the Caspase pathway and reverses EMT; Dictamni, Kochia, frankincense, myrrh, alum, etc.: regulate mucosal immunity, anti-inflammation, and rebuild the microecology; each component works synergistically, adopting the mechanism of local epithelial microecological repair + mucosal immunity, and synergistically fights HPV at multiple targets.

[0015] Preferably, the proportion of each component of the aqueous gel carrier, in percentage, is such that the total content of glycerol, propylene glycol, carbomer, and alginate is no more than 8%, and the balance is purified water.

[0016] Preferably, the extraction method of each plant component extract is at least one of water decoction, alcohol precipitation, reflux extraction, ultrasonic extraction, enzymatic extraction, membrane separation, macroporous resin adsorption or supercritical extraction.

[0017] Preferably, it is used for the persistent positive state after cervical HPV infection.

[0018] The present invention also provides a method for preparing the anti-HPV gel preparation, comprising the following steps:

[0019] 1) Mixing the extracts of rhubarb, scutellaria, coptis root, phellodendron, sophora flavescens, cnidium monnieri, alum, gallnut, radix chinensis, angelica dahurica, bark of Dictamni, kochia scoparia, frankincense, and myrrh in proportion by mass, stirring evenly to obtain a compound plant extract;

[0020] 2) Prepare an aqueous gel carrier by mixing glycerol, propylene glycol, carbomer, alginate, and purified water, and let it stand for a while to mature before using as the gel carrier;

[0021] 3) Mixing the compound plant extract and the aqueous gel carrier according to a certain ratio, stirring them evenly using a stirring device to form a stable traditional Chinese medicine gel preparation system;

[0022] 4) Filling the prepared traditional Chinese medicine gel into vaginal external propellers in 3-gram or 5-gram quantities, sterilizing, and finally packaging and storing to obtain a plant compound anti-HPV gel preparation for gynecological use.

[0023] Preferably, the mass percentage of the compound plant extract in the anti-HPV gel preparation is 8-15%, and the pH of the anti-HPV gel preparation is 4.72.

[0024] Furthermore, the compound plant extract and the aqueous gel carrier are composed in a mass ratio of 1:9, and the compound plant extract accounts for 10% of the total mass of the gel.

[0025] Beneficial effects:

[0026] 1) Block viral integration and replication, inhibit E6 / E7 oncogenic protein expression, disrupt L1 capsid assembly, cover key nodes in the viral life cycle, and synergize multiple targets against HPV to reduce the risk of escape;

[0027] 2) Reduce the risk of cervical lesion progression by inhibiting the NF-κB / EMT signaling axis;

[0028] 3) Enhance local mucosal immune factors (such as sIgA and IFN-β), restore bacterial flora homeostasis, regulate immunity and microenvironment simultaneously, and improve HPV clearance efficiency;

[0029] 4) Achieve safe and effective local intervention through viral suppression, oncogenic protein regulation, and tissue repair;

[0030] 5) No hormones or antibiotics, avoiding the side effects of traditional treatments and improving patient compliance;

[0031] 6) Covering multiple links such as viral replication, transcriptional regulation, and immune regulation, it avoids the limitations of a single target and has a complete mechanism;

[0032] 7) It is non-irritating, its pH matches the vaginal environment, it is highly safe, and it takes into account both antiviral and microenvironment repair, making it suitable for long-term intervention.

[0033] In summary, the present invention provides an anti-HPV gel preparation for local use and multi-target intervention based on active ingredients of botanicals, for female populations who persist in HPV infection but have not yet developed advanced cervical lesions. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 Heat map of changes in HPV viral load in patients;

[0035] Figure 2 This is a comparison chart of the efficacy of negative conversion in two groups of people infected with high-risk and low-risk HPV. DETAILED DESCRIPTION

[0036] Experimental methods:

[0037] ⒜ (HPV16) inactivation test

[0038] Sample information: Antiviral gel stock solution was used as the test substance;

[0039] Virus name: HPV pseudovirus (HPV16-GFP); Source: Beijing Yunling Biotechnology;

[0040] Host cell: 293FT

[0041] Neutralizer formula: D / E Neutralizing Broth;

[0042] Culture medium: DMEM / 2891229, FBS / 2582168P;

[0043] Instruments and equipment: integrated fluorescence microscopy system light source BZ-X800LE, microscope BZ-X810 (WPE-TW0098), CO2 incubator BPN-CRH (WPE-RH0274), inverted microscope LWD300-38LT (WPE-RH0164), biological safety cabinet BSC-1604ⅡB2 (WPE-RH0313);

[0044] Operation steps: First, 293FT cells are inoculated into a 96-well cell culture plate and the cells are attached to the wall for later use; then 10 μL of virus solution is added to the 10mm×10mm sample and control sample, spread evenly, and act for 12 hours. Then, it is transferred to 1 mL of neutralizer or culture medium, vortexed to mix, and neutralized for 10 minutes as the first gradient, and then 10-fold gradient dilution is performed in sequence. Take 100 μL of gradient diluted virus solution and add it to the cells. Set 4 wells for detection for each concentration gradient. Culture in a carbon dioxide incubator at 37°C for 72 hours, and then measure the residual virus titer. Use a fluorescence microscope to count the number of fluorescent cells and calculate the FFU (fluorescence forming unit) of the virus.

[0045] The experiment was conducted with reference to 2.1.1.10 of the Technical Specifications for Disinfection (2002 edition);

[0046] Testing environment: temperature: 22.3°C, humidity: 50%RH.

[0047] ⒝Multiple complete skin irritation tests

[0048] Animal species: Japanese large-eared white rabbit; Grade: Ordinary; Number: 3; Gender: Female; Weight: Initial weight: 2.16 kg–2.20 kg; Source: Tianjin Yuda Experimental Animal Breeding Co., Ltd. (License No.: SCXK(Jin)2021-0001), Certificate No.: 120906251100020962;

[0049] Experimental conditions: Normal environment, temperature 22.2℃-23.4℃, relative humidity 49.7%-59.6%, 12-h light-dark cycle, experimental animal use license number SYXK(Liaoning)2024-0001;

[0050] Instruments and equipment: electronic platform scale / TCS-TH-226-200 / YQ4004;

[0051] Exposure method: Follow the standard exposure procedure and use the test substance for exposure. 24 hours before exposure, remove the hair on both sides of the spine of the Japanese white rabbit's back with electric clippers, covering an area of ​​approximately 3cm×3cm. The next day, apply 0.5mL of the test substance to one side of the intact skin of 2.5cm×2.5cm, and apply purified water to the other side as a control. Rinse with purified water 4 hours after application to remove any residue. Apply the test substance once a day for 14 consecutive days.

[0052] Animal observation: Observe the back of the animal with the naked eye 24 hours after each sample application, and score and record the skin irritation reaction according to the requirements of the specification. The observation period is 14 days.

[0053] ⒞Multiple vaginal mucosal irritation test

[0054] Experimental animals and breeding environment

[0055] Animal strain: Japanese large-eared white rabbit, grade: ordinary, sex: female, weight: 2.16 kg–2.20 kg; number: 6; animal source: Tianjin Yuda Experimental Animal Breeding Co., Ltd., experimental animal production license number: SCXK(Jin)2021-0001; experimental animal quality certificate number: 120906251100020962

[0056] Experimental conditions: Ordinary environment, temperature: 22.6℃~23.0℃, relative humidity: 56.4%~59.6%, light intensity: 12h light-dark alternation, experimental animal use license number: SYXK(Liaoning)2024-0001;

[0057] Instruments and equipment:

[0058] Electronic platform scale / TCS-TH-226-200 / YQ4004, biological microscope / SK160+BTX10 / YQ4014, biological tissue slice baking machine IYD-AB2 / YQ4007, tissue dehydrator / YD-12PYQ4008, tissue stainer / YD-700 / YQ4011, tissue slicer / YD-335 / YQ4009, embedding machine / YD-6L / YQ4010;

[0059] Animals were divided into two groups: the poisoned group and the negative control group, with 3 animals in each group.

[0060] Exposure method: Follow the standard exposure procedure and use the test substance for exposure. First, fix the experimental animal on its back to expose the perineum and vaginal opening. Fill a 2mL syringe with the test substance and gently insert it into the vagina for about 4-5cm. Slowly inject 2mL of the test substance into each animal. Remove the soft tube to complete the exposure. Repeat the exposure every 24 hours for 5 consecutive days. The experimental animals in the control group receive normal saline and are treated in the same manner as the exposure group.

[0061] Sampling and slide preparation: 24 hours after the last exposure, animals were sacrificed by gas embolism. The intact vagina was removed by laparotomy and longitudinally incised to observe for congestion and edema. The vagina was then fixed in 10% formalin for 24 hours. Tissue slides were prepared from the two ends and three central locations of the vagina and stained with hematoxylin and eosin for histopathological examination.

[0062] Observation: All slide specimens were individually numbered. The irritation response scores of the three sites of the three animals in the dose group were added together and then divided by the total number of observations (number of animals × 3) to obtain the average score of the vaginal mucosal irritation response of the dose group and record it. The scoring method of the control group was the same and recorded.

[0063] Example 1:

[0064] The following fourteen medicinal materials were extracted by decoction and mixed in proportion: Rheum officinale 13.89%, Scutellaria baicalensis 13.89%, Coptis chinensis 13.89%, Phellodendron amurense 13.89%, Sophora flavescens 13.89%, Cnidium monnieri 2.78%, Alum 2.78%, Gallnut chinensis 5.56%, Trichosanthes kirilowii 4.17%, Angelica dahurica 4.17%, Dictamni cortex 2.78%, Kochia scoparia 2.78%, Frankincense 2.78%, Myrrh 2.78%;

[0065] The proportions of the components of the aqueous gel carrier, in terms of percentage, are as follows: the total content of glycerol, propylene glycol, carbomer, and alginate is 8%, and purified water is 92%.

[0066] Prepare according to the following steps:

[0067] 1) Mixing the extracts of rhubarb, scutellaria, coptis root, phellodendron, sophora flavescens, cnidium monnieri, alum, gallnut, radix chinensis, angelica dahurica, bark of Dictamni, kochia scoparia, frankincense, and myrrh in proportion by mass, stirring evenly to obtain a compound plant extract;

[0068] 2) Prepare an aqueous gel carrier by mixing glycerol, propylene glycol, carbomer, alginate, and purified water, and let it stand for a while to mature before using as the gel carrier;

[0069] 3) Mixing the compound plant extract and the aqueous gel carrier in a mass ratio of 1:9, and stirring evenly using a stirring device to form a stable traditional Chinese medicine gel preparation system;

[0070] 4) Filling the prepared traditional Chinese medicine gel into a vaginal external propeller in a volume of 3 grams, sterilizing, and finally packaging and storing to obtain a plant compound anti-HPV gel preparation for gynecological use.

[0071] Example 2:

[0072] The following fourteen medicinal materials were extracted by alcohol precipitation, reflux extraction, ultrasonic extraction, enzymatic extraction, membrane separation, macroporous resin adsorption or supercritical extraction, and mixed in proportion: Rheum officinale 13.89%, Scutellaria baicalensis 13.89%, Coptis chinensis 13.89%, Phellodendron amurense 13.89%, Sophora flavescens 13.89%, Cnidium monnieri 2.78%, Alum 2.78%, Galla chinensis 5.56%, Ligusticum chuanxiong 4.17%, Angelica dahurica 4.17%, Dictamni cortex 2.78%, Kochia scoparia 2.78%, Frankincense 2.78%, Myrrh 2.78%;

[0073] The proportions of the components of the aqueous gel carrier, in terms of percentage, are as follows: the total content of glycerol, propylene glycol, carbomer, and alginate is 7%, and purified water is 93%.

[0074] Prepare according to the following steps:

[0075] 1) Mixing the extracts of rhubarb, scutellaria, coptis root, phellodendron, sophora flavescens, cnidium monnieri, alum, gallnut, radix chinensis, angelica dahurica, bark of Dictamni, kochia scoparia, frankincense, and myrrh in proportion by mass, stirring evenly to obtain a compound plant extract;

[0076] 2) Prepare an aqueous gel carrier by mixing glycerol, propylene glycol, carbomer, alginate, and purified water, and let it stand for a while to mature before using as the gel carrier;

[0077] 3) Mixing the compound plant extract and the aqueous gel carrier according to a certain ratio, stirring them evenly using a stirring device to form a stable traditional Chinese medicine gel preparation system;

[0078] 4) Filling the prepared traditional Chinese medicine gel into a vaginal external propeller in a volume of 3 grams, sterilizing, and finally packaging and storing to obtain a plant compound anti-HPV gel preparation for gynecological use.

[0079] The mass percentage of the compound plant extract in the gel preparation is 8%.

[0080] Example 3:

[0081] The following fourteen medicinal materials were extracted by reflux extraction, ultrasonic extraction, enzymatic extraction, membrane separation, macroporous resin adsorption or supercritical extraction, and mixed in proportion: Rheum officinale 13.89%, Scutellaria baicalensis 13.89%, Coptis chinensis 13.89%, Phellodendron amurense 13.89%, Sophora flavescens 13.89%, Cnidium monnieri 2.78%, Alum 2.78%, Galla chinensis 5.56%, Glehnia littoralis 4.17%, Angelica dahurica 4.17%, Dictamnus divaricata 2.78%, Kochia scoparia 2.78%, Frankincense 2.78%, Myrrh 2.78%;

[0082] The proportions of the components of the aqueous gel carrier, in terms of percentage, are as follows: the total content of glycerol, propylene glycol, carbomer, and alginate is 6%, and purified water is 94%.

[0083] Prepare according to the following steps:

[0084] 1) Mixing the extracts of rhubarb, scutellaria, coptis root, phellodendron, sophora flavescens, cnidium monnieri, alum, gallnut, radix chinensis, angelica dahurica, bark of Dictamni, kochia scoparia, frankincense, and myrrh in proportion by mass, stirring evenly to obtain a compound plant extract;

[0085] 2) Prepare an aqueous gel carrier by mixing glycerol, propylene glycol, carbomer, alginate, and purified water, and let it stand for a while to mature before using as the gel carrier;

[0086] 3) Mixing the compound plant extract and the aqueous gel carrier according to a certain ratio, stirring them evenly using a stirring device to form a stable traditional Chinese medicine gel preparation system;

[0087] 4) Filling the prepared traditional Chinese medicine gel into vaginal external propellers in a 5-gram package, sterilizing, and finally packaging and storing to obtain a plant compound anti-HPV gel preparation for gynecological use.

[0088] The mass percentage of the compound plant extract in the gel preparation is 12%.

[0089] Example 4:

[0090] The following 14 medicinal materials were extracted by ultrasonic extraction, enzymatic extraction, membrane separation, macroporous resin adsorption or supercritical extraction, and mixed in proportion: Rheum officinale 13.89%, Scutellaria baicalensis 13.89%, Coptis chinensis 13.89%, Phellodendron amurense 13.89%, Sophora flavescens 13.89%, Cnidium monnieri 2.78%, Alum 2.78%, Galla chinensis 5.56%, Glehnia littoralis 4.17%, Angelica dahurica 4.17%, Dictamni cortex 2.78%, Kochia scoparia 2.78%, Frankincense 2.78%, Myrrh 2.78%;

[0091] The proportions of the components of the aqueous gel carrier, in terms of percentage, are as follows: the total content of glycerol, propylene glycol, carbomer, and alginate is 9%, and purified water is 91%.

[0092] Prepare according to the following steps:

[0093] 1) Mixing the extracts of rhubarb, scutellaria, coptis root, phellodendron, sophora flavescens, cnidium monnieri, alum, gallnut, radix chinensis, angelica dahurica, bark of Dictamni, kochia scoparia, frankincense, and myrrh in proportion by mass, stirring evenly to obtain a compound plant extract;

[0094] 2) Prepare an aqueous gel carrier by mixing glycerol, propylene glycol, carbomer, alginate, and purified water, and let it stand for a while to mature before using as the gel carrier;

[0095] 3) Mixing the compound plant extract and the aqueous gel carrier according to a certain ratio, stirring them evenly using a stirring device to form a stable traditional Chinese medicine gel preparation system;

[0096] 4) Filling the prepared traditional Chinese medicine gel into vaginal external propellers in a 5-gram package, sterilizing, and finally packaging and storing to obtain a plant compound anti-HPV gel preparation for gynecological use.

[0097] The mass percentage of the compound plant extract in the gel preparation is 14%.

[0098] Example 5:

[0099] The following 14 medicinal materials were extracted by enzymatic extraction method and mixed in proportion: Rheum officinale 13.89%, Scutellaria baicalensis 13.89%, Coptis chinensis 13.89%, Phellodendron amurense 13.89%, Sophora flavescens 13.89%, Cnidium monnieri 2.78%, Alum 2.78%, Gallnut chinensis 5.56%, Glehnia littoralis 4.17%, Angelica dahurica 4.17%, Dictamni bark 2.78%, Kochia scoparia 2.78%, Frankincense 2.78%, Myrrh 2.78%;

[0100] The proportions of the components of the aqueous gel carrier, calculated in percentage, are as follows: the total content of glycerol, propylene glycol, carbomer, and alginate is 10%, and purified water is 90%.

[0101] Prepare according to the following steps:

[0102] 1) Mixing the extracts of rhubarb, scutellaria, coptis root, phellodendron, sophora flavescens, cnidium monnieri, alum, gallnut, radix chinensis, angelica dahurica, bark of Dictamni, kochia scoparia, frankincense, and myrrh in proportion by mass, stirring evenly to obtain a compound plant extract;

[0103] 2) Prepare an aqueous gel carrier by mixing glycerol, propylene glycol, carbomer, alginate, and purified water, and let it stand for a while to mature before using as the gel carrier;

[0104] 3) Mixing the compound plant extract and the aqueous gel carrier according to a certain ratio, stirring them evenly using a stirring device to form a stable traditional Chinese medicine gel preparation system;

[0105] 4) Filling the prepared traditional Chinese medicine gel into vaginal external propellers in a 5-gram package, sterilizing, and finally packaging and storing to obtain a plant compound anti-HPV gel preparation for gynecological use.

[0106] The mass percentage of the compound plant extract in the gel preparation is 15%.

[0107] Example 6:

[0108] The plant component extraction method in Example 1 was changed to a membrane separation method, and the rest remained unchanged.

[0109] Example 7:

[0110] The plant component extraction method in Example 2 was changed to a macroporous resin adsorption method, and the rest remained unchanged.

[0111] Example 8:

[0112] The plant component extraction method in Example 3 was changed to supercritical extraction, and the rest remained unchanged.

[0113] Experiments and data:

[0114] 1. The anti-HPV gel preparation in Example 1 was tested. The results of the HPV16 pseudovirus inactivation test were as follows:

[0115]

[0116] After testing, the sample's logarithmic inactivation value for human papillomavirus (HPV16) after 12 hours of action was 1.08, and the virus inactivation rate was 91.73%.

[0117] 2. The anti-HPV gel preparation in Example 8 was tested. The results of the skin local reactions of three Japanese white rabbits during the multiple intact skin irritation tests are shown in the following table:

[0118]

[0119] After testing, the sample's irritation index to test animals was 0.00, and the skin irritation intensity was non-irritating.

[0120] 3. The anti-HPV gel preparation described in Example 4 was tested. The test sample was used directly. After exposure, the vagina was removed and visual inspection revealed no vascular congestion or edema in either the control or exposure groups. The vaginal mucosal irritation response was scored based on histopathological examination results. The results are shown in the following table:

[0121]

[0122] After testing, the sample's multiple vaginal mucosal irritation index to Japanese large-eared white rabbits was <1, and the irritation intensity was non-irritating.

[0123] Use Cases:

[0124] Clinical observation data show that the multi-target intervention plant compound gynecological anti-HPV gel preparation of the present invention was used to treat more than 200 women with persistent high-risk HPV infection. The patients were all characterized by persistent HPV infection that had not turned negative within one year. After continuous medication for 3-6 months, the HPV negative conversion rate reached 86%.

[0125] For those infected with low-risk HPV, local treatment with a multi-target intervention plant-based compound gynecological anti-HPV gel preparation can be effectively used for the intervention and removal of HPV warts on the vulva and cervix, with a negative conversion rate of 90.2%.

[0126] Treatment period: 3-6 months;

[0127] Treatment: After microwave treatment of the cervix, apply traditional Chinese medicine once daily, using Formula 1 and Formula 2 for the first 15 days, followed by Formula 2 thereafter. For patients with a predominantly abnormal constitution, oral traditional Chinese medicine should be added to improve their constitution and immune system. Monitor HPV viral load every 2-3 months to determine whether to proceed with the next treatment cycle.

[0128] Typical cases:

[0129] Case 1: Patient Ma, female, 59 years old, visited the hospital on August 19, 2024;

[0130] Chief complaint: Menopause for 7 years, persistent infection with HPV51, 59, 42, and 43 for more than four years, and long-term poor sleep.

[0131] Current medical history: 7 years of menopause. In October 2020, low-risk cervical HPV infection was discovered, and interferon vaginal application was administered. On February 5, 2021, cervical examination was positive for HPV types 51, 59, 42, 43, and 44. Gongyikang vaginal application was administered. On February 10, a colposcopy was performed, suggesting chronic cervicitis and endocervicitis. On August 10, 2021, a cervical TCT examination revealed atypical squamous epithelium and high-risk HPV. In March 2022, HPV types 42, 43, 51, and 59 were positive. TCT DNA showed a small number of DNA-diploid cells. Biopsy pathology revealed chronic inflammation of the cervix (at 3, 6, 9, and 12 o'clock), with focal LSIL suspected. On March 15, 2022, she underwent a 1-year cervical, vulvar, and perianal endoscopic procedure. Interferon 2b suppositories and Baofukang suppositories were applied alternately vaginally. A follow-up examination in October 2022 revealed positive results for HPV types 42, 43, and 59. TCT DNA showed a small number of DNA-positive cells (ASCUS). A follow-up TCT examination in May 2023 revealed ASC-US and positive results for HPV types 42, 43, 51, and 59. Endocervical scraping revealed very few endocervical cells. Following the initial diagnosis and treatment, a follow-up TCT DNA examination on July 10, 2024, revealed ASC-US and positive results for HPV types 42, 43, 51, and 59. The patient requested traditional Chinese medicine treatment.

[0132] Traditional Chinese Medicine: Waking easily from sleep, waking between 11am and 1am every night, difficulty falling asleep after waking, poor sleep quality, occasional grogginess in the morning, incomplete bowel movements, pale red tongue with a thin white coating, and a deep, thready pulse. Traditional Chinese Medicine diagnosis: Spleen and Kidney Yang deficiency, and gallbladder deficiency.

[0133] Treatment: 1 local cervical treatment + 3 months of Chinese medicine. On April 17, 2025, HPV43 and 42 turned negative, HPV51 and 59 were positive but the viral load decreased. TCT: ASCUS. Now the sleep has improved and the condition is good. Further treatment is in progress. For details, see Figure 1

[0134] Case 2: Patient Li, a 45-year-old female, presented with a cervical HPV test that was positive for HPV51 and 52, but negative for TCT. She also had visible cervical polypoid hyperplasia and adenomyosis at the cervical ostium. Treatment included one topical cervical treatment followed by three months of Chinese herbal remedies. The patient's HPV status turned negative, and the cervical polyps resolved.

[0135] Moreover, during the entire use process, patients tolerated it well and no adverse reactions were observed. Therefore, the gel preparation of the present invention, as an innovative external-use gel preparation of Chinese herbal extracts, not only improves the effectiveness of treating persistent high-risk HPV infection, but also takes into account patient comfort and compliance, and has good market conversion prospects.

[0136] The content of the above embodiments is only a specific interpretation of the technical solution of the present invention, which does not mean a limitation of the technical solution of the present invention. After seeing the content of the present invention, those skilled in the art can still obtain technical solutions by simply adding, subtracting or adjusting the above-mentioned solutions within the technical protection scope of the present invention.

Claims

1. A plant compound anti-HPV gel preparation for gynecological use based on multi-target intervention, consisting of a compound plant extract and an aqueous gel carrier, characterized in that: The compound plant extract is derived from: rhubarb, scutellaria, coptis root, phellodendron, sophora flavescens, cnidium monnieri, alum, gallnut, scutellaria baicalensis, angelica dahurica, dandelion bark, kochia scoparia, frankincense and myrrh; The aqueous gel carrier consists of glycerol, purified water, propylene glycol, carbomer and alginate.

2. The plant compound anti-HPV gel preparation for gynecology based on multi-target intervention according to claim 1, characterized in that: The ratio of the components of the compound plant extract is: m Cnidium monnieri: m Alum: m Dictamnus cortex: m Kochia scoparia: m Frankincense: m Myrrh: m Gallnut = 1:1:1:1:1:1:2; m Gastrodia elata: m Angelica dahurica = 1:1; m Rhubarb: m Scutellaria baicalensis: m Coptis chinensis: m Phellodendron amurense: m Sophora flavescens = 1:1:1:1:1:

1.

3. The plant compound anti-HPV gel preparation for gynecology based on multi-target intervention according to claim 2, characterized in that: m Dictamnus cortex: m Galla chinensis: m Angelica dahurica: m Scutellaria baicalensis = 1:2:1.5:5.

0.

4. The plant compound anti-HPV gel preparation for gynecology based on multi-target intervention according to claim 1, characterized in that: The proportion of each component of the aqueous gel carrier, calculated in percentage, is such that the total content of glycerol, propylene glycol, carbomer, and alginate is no more than 8%, and the balance is purified water.

5. The plant compound anti-HPV gel preparation for gynecology based on multi-target intervention according to claim 1, characterized in that: The extraction method of each plant component extract is at least one of water decoction, alcohol precipitation, reflux extraction, ultrasonic extraction, enzymatic extraction, membrane separation, macroporous resin adsorption or supercritical extraction.

6. The method for preparing a botanical compound gynecological anti-HPV gel preparation based on multi-target intervention according to any one of claims 1 to 5, characterized in that: The steps include: 1) Mixing the extracts of rhubarb, scutellaria, coptis root, phellodendron, sophora flavescens, cnidium monnieri, alum, gallnut, radix chinensis, angelica dahurica, bark of Dictamni, kochia scoparia, frankincense, and myrrh in proportion by mass, stirring evenly to obtain a compound plant extract; 2) Prepare an aqueous gel carrier by mixing glycerol, propylene glycol, carbomer, alginate, and purified water, and let it stand for a while to mature before using as the gel carrier; 3) Mixing the compound plant extract and the aqueous gel carrier according to a certain ratio, stirring them evenly using a stirring device to form a stable traditional Chinese medicine gel preparation system; 4) Filling the prepared traditional Chinese medicine gel into vaginal external propellers in 3-gram or 5-gram quantities, sterilizing, and finally packaging and storing to obtain a plant compound anti-HPV gel preparation for gynecological use.

7. The preparation method according to claim 6, characterized in that The mass percentage of the compound plant extract in the anti-HPV gel preparation is 8-15%.

8. The plant compound anti-HPV gel preparation for gynecology based on multi-target intervention according to claim 1, characterized in that: Used for the persistent positive state after cervical HPV infection.

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