Strychnos nuxvomica for intervening in rheumatoid arthritis as well as processing method and application thereof

Abstract

The invention discloses nux vomica for intervening in rheumatoid arthritis and a processing method and application thereof, and belongs to the technical field of traditional Chinese medicinal material processing. The processing method comprises the following steps: decocting crude semen strychni with a water decoction of rhizoma atractylodis macrocephalae, filtering, and drying filter residues to obtain the processed semen strychni. According to the processing method, the medicine effect and safety of the semen strychni are remarkably improved through the unique processing technology that the rhizoma atractylodis macrocephalae water decoction is adopted to decoct the semen strychni crude product, the original anti-inflammatory, analgesic and immunoregulation activity of the rhizoma atractylodis macrocephalae and the semen strychni crude product are reserved, the synergistic interaction effect is generated, the toxicity of the semen strychni is systematically reduced, and the effect is remarkable. The pharmaceutical safety is greatly improved while the treatment effect is enhanced, and the pharmaceutical composition has important clinical application value and market prospect.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine processing technology, and in particular to a method of processing and application of Strychnos nux-vomica for intervening in rheumatoid arthritis. Background Technology

[0002] Rheumatoid arthritis (RA) is a chronic autoimmune inflammatory disease characterized by morning stiffness, joint pain, swelling, and tenderness. These symptoms primarily manifest as symmetrical polyarthritis, mainly affecting small joints in the hands and feet, such as the wrists and metacarpophalangeal joints. The disease is characterized by chronic inflammation of the synovial membrane and erosive destruction of the joints, which can lead to joint deformities or loss of function in severe cases. Patients typically experience morning stiffness, characterized by joint pain upon waking, which is relieved by activity and usually lasts for more than half an hour. Currently, RA treatment mainly involves anti-inflammatory analgesics, immunomodulatory agents, and biologics. However, long-term use can easily lead to drug resistance and adverse reactions. Therefore, exploring highly effective and low-toxicity traditional Chinese medicine interventions is of significant clinical importance.

[0003] Strychnos nux-vomica L., a traditional Chinese medicine, has the effects of clearing the meridians, relieving pain, dispersing nodules, and reducing swelling. It is often used to treat stubborn rheumatism, traumatic injuries, carbuncles, and boils. Its pharmacologically active components (such as alkaloids and iridoid glycosides) have significant analgesic, anti-inflammatory, cartilage repair-promoting, and immunomodulatory effects, which meet the treatment needs of rheumatoid arthritis (RA). However, raw Strychnos nux-vomica is highly toxic and can only be used externally. Although processing can reduce its toxicity for internal use, the sand-frying method specified in the current Chinese Pharmacopoeia (2025 edition) relies on empirical control, resulting in unstable toxicity reduction and efficacy. Furthermore, the retention of effective components such as alkaloids and the balance of toxicity are difficult to precisely control, which restricts its clinical application.

[0004] Existing research indicates that the combination of Strychnos nux-vomica and Atractylodes macrocephala can reduce toxicity through mechanisms such as decreasing alkaloid content and inhibiting the absorption of toxic components, suggesting that excipient processing is an effective way to optimize its safety. However, traditional processing methods (such as processing with licorice or children's urine) are complex and have low standardization, making it difficult to meet the needs of modern industrial production. Therefore, developing a Strychnos nux-vomica processing method based on a toxicity-reduction and efficacy-enhancing mechanism with controllable processes is of great significance for improving its applicability in the treatment of rheumatoid arthritis (RA).

[0005] In summary, considering the pathological characteristics of RA and the pharmacological advantages of strychnine, there is an urgent need to establish a scientific and standardized processing technique that can systematically reduce toxicity while preserving anti-inflammatory, analgesic, and immunomodulatory activities, thus providing a safer and more effective traditional Chinese medicine intervention strategy for rheumatoid arthritis. Summary of the Invention

[0006] The purpose of this invention is to provide a method for processing and applying Strychnos nux-vomica (a type of strychnine) to intervene in rheumatoid arthritis, thereby addressing the problems existing in the prior art. This invention utilizes a unique processing technique involving decocting raw Strychnos nux-vomica with Atractylodes macrocephala decoction, significantly enhancing the efficacy and safety of Strychnos nux-vomica. The processing method of this invention not only preserves the original anti-inflammatory, analgesic, and immunomodulatory activities of both ingredients but also produces a synergistic effect, systematically reducing the toxicity of Strychnos nux-vomica. While enhancing therapeutic effects, it also significantly improves medication safety, possessing significant clinical application value and market prospects.

[0007] To achieve the above objectives, the present invention provides the following solution:

[0008] This invention provides a method for processing nux vomica for intervention in rheumatoid arthritis, comprising decocting raw nux vomica in an aqueous decoction of Atractylodes macrocephala, filtering, and drying the residue to obtain processed nux vomica.

[0009] Furthermore, the method for preparing the decoction of Atractylodes macrocephala includes the following steps: soaking Atractylodes macrocephala in water and then decocting it for the first time, filtering it to obtain the first filtrate; adding water to the residue again and decocting it for the second time, filtering it to obtain the second filtrate, and combining the two filtrates to obtain the decoction of Atractylodes macrocephala.

[0010] Furthermore, during the first decoction, the ratio of Atractylodes macrocephala to water is 1g:20mL; the first decoction is brought to a boil over high heat and then simmered over low heat for 30 minutes.

[0011] Furthermore, during the second decoction, the ratio of the filter residue to water is 1g:10mL; the second decoction is to bring to a boil over high heat, then simmer over low heat for 20 minutes.

[0012] Furthermore, the raw nux vomica and the aqueous decoction of Atractylodes macrocephala are mixed at a ratio of 1g:60mL.

[0013] Furthermore, the soaking time is 30 minutes.

[0014] Furthermore, the decoction is prepared by bringing the mixture to a boil over high heat and then simmering it over low heat for 1 hour.

[0015] Furthermore, the drying process involves drying at 60°C for 4 hours.

[0016] The present invention also provides a processed strychnine obtained according to the described strychnine processing method.

[0017] The present invention also provides the application of the processed Strychnos nux-vomica in the preparation of a drug for treating rheumatoid arthritis.

[0018] The present invention discloses the following technical effects:

[0019] The method for processing nux vomica provided by this invention, through a unique processing technique of decocting raw nux vomica with Atractylodes macrocephala decoction, significantly enhances the efficacy and safety of nux vomica. Experiments have confirmed that the resulting Atractylodes macrocephala-processed nux vomica slices are significantly more effective in treating rheumatoid arthritis than the decoction of raw nux vomica and the decoction of raw nux vomica and Atractylodes macrocephala. It not only retains the original anti-inflammatory, analgesic, and immunomodulatory activities of both ingredients but also produces a synergistic effect, providing a more efficient traditional Chinese medicine intervention for clinical practice.

[0020] Furthermore, the processing method of this invention, through the optimization of the compatibility of Atractylodes macrocephala and the processing technology, systematically reduces the toxicity of Strychnos nux-vomica, significantly improving medication safety while enhancing therapeutic efficacy. This invention provides an innovative traditional Chinese medicine decoction piece for the treatment of rheumatoid arthritis, possessing both significant therapeutic efficacy and low toxicity, and has important clinical application value and market prospects. Attached Figure Description

[0021] 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.

[0022] Figure 1 The effect of different processed forms of Strychnos nux-vomica on the spleen index (A) and thymus index (B) of RA rats;

[0023] Figure 2 Phenotypic effect of different processed forms of Strychnos nux-vomica on the appearance of paw swelling in RA rats;

[0024] Figure 3 A statistical graph showing the effect of different processed forms of Strychnos nux-vomica on paw swelling in rats with rhabdomyorrhea. Detailed Implementation

[0025] 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.

[0026] 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. Any stated value or intermediate value within a stated range, as well as each smaller range between 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.

[0027] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of this invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials related to the documents. In case of conflict with any incorporated document, the content of this specification shall prevail.

[0028] Without departing from the scope or spirit of the present invention, various modifications and variations can be made to the specific embodiments of the specification of the present invention, which will be obvious to those skilled in the art. Other embodiments obtained from the specification of the present invention will be obvious to those skilled in the art. The specification and examples of the present invention are merely exemplary.

[0029] As used herein, "comprising", "including", "having", "containing", and the like are open-ended terms, meaning including but not limited to.

[0030] The animals used in the embodiments of the present invention were 60 female SD rats with a body weight in the range of (180 ± 20) g, provided by the Experimental Center of Jiangxi University of Traditional Chinese Medicine, with the certificate number SCXK(Gan)2023-0001. The room temperature was controlled within 20 - 23°C, and the relative humidity was 45% - 55%. After 1 week of adaptive feeding, a rat model of adjuvant arthritis induced by complete Freund's adjuvant was constructed.

[0031] The medicinal materials used in the embodiments of the present invention: Strychnos seeds were purchased from Jiangxi Ancient Han Refined Traditional Chinese Medicine Pieces Co., Ltd., and Atractylodes rhizomes were purchased from Tiantai, Zhejiang. They were respectively identified by Professor Gong Qianfeng of Jiangxi University of Traditional Chinese Medicine as the dried ripe seeds of Strychnos nux-vomica L., a plant of the Loganiaceae family, with the batch number 19042613; and the dried rhizomes of Atractylodes macrocephala Koidz., a plant of the Asteraceae family, with the batch number 20211214.

[0032] The reagents used in the embodiments of this invention: tumor necrosis factor-α (TNF-α) detection kit (catalog number: MM-0180R1), interleukin-6 (IL-6) detection kit (catalog number: MM-0190R1), and IL-1β detection kit (catalog number: MM-0047R1) were all purchased from Ruixin Co., Ltd.; the water was Watson distilled water. The alanine aminotransferase (ALT) assay kit (batch number: 24062519-1), aspartate aminotransferase (AST) assay kit (batch number: 24051020-2), blood urea nitrogen (BUN) assay kit (batch number: 24060628-1), and creatinine (CRE) assay kit (batch number: 24080527-1) were all purchased from Wuhan Shengzhiyuan Biotechnology Co., Ltd. Unless otherwise specified, other materials and reagents were obtained through routine purchases.

[0033] Example

[0034] I. Experimental Methods

[0035] 1. Preparation of different processed strychnine slices

[0036] Preparation of raw nux vomica: Soak nux vomica in water for 2 days, moisten for half a day, remove the skin and hair, cut into 2mm thick slices, and dry at 60℃ for 4 hours to obtain raw nux vomica slices.

[0037] Preparation of Strychnos nux-vomica processed with Atractylodes macrocephala:

[0038] (1) Take an appropriate amount of Atractylodes macrocephala, add water and soak for 30 minutes at a ratio of 1g:20mL, bring to a boil over high heat and then simmer over low heat for 30 minutes, filter, add water to the residue at a ratio of 1g:10mL, bring to a boil over high heat and then simmer over low heat for another 20 minutes, filter, combine the two filtrates, and prepare a 1g / 10mL Atractylodes macrocephala decoction for later use.

[0039] (2) Take an appropriate amount of nux vomica, add Atractylodes macrocephala decoction at a ratio of 1g:60mL, bring to a boil over high heat, then simmer over low heat for 1 hour, filter, and dry the residue at 60℃ for 4 hours to obtain processed nux vomica slices.

[0040] 2. Preparation of decoction

[0041] Preparation of raw nux vomica decoction: Take an appropriate amount of raw nux vomica, soak it in distilled water at a ratio of 1g:130mL for 30 minutes, then heat to a boil. After boiling over high heat, simmer over low heat for 30 minutes, stirring. Filter while hot. The residue is then boiled again in distilled water at a ratio of 1g:80mL for 20 minutes. Filter and combine the filtrates to obtain raw nux vomica decoction for later use.

[0042] Preparation of Atractylodes macrocephala + Strychnos nux-vomica decoction: Strychnos nux-vomica and Atractylodes macrocephala were prepared at a mass ratio of 1:6, that is, 1g of Strychnos nux-vomica and 6g of Atractylodes macrocephala were accurately weighed, soaked in 400mL of distilled water for 30min, and then heated to a boil. Initially, high heat was used, and after boiling, low heat was used to boil for 30min. Stirring was carried out, and the mixture was filtered while hot. The residue was then boiled again with distilled water at a material-liquid ratio of 1g:35mL for 20min. The mixture was filtered, and the filtrates were combined to obtain the Atractylodes macrocephala + Strychnos nux-vomica decoction.

[0043] Preparation of Atractylodes macrocephala-processed Strychnos nux-vomica decoction: Take an appropriate amount of Atractylodes macrocephala-processed Strychnos nux-vomica, soak it in distilled water at a ratio of 1g:130mL for 30 minutes, then heat to a boil. After boiling over high heat, simmer over low heat for 30 minutes, stirring, and filter while hot. The residue is then boiled again in distilled water at a ratio of 1g:80mL for 20 minutes, filtered, and the filtrates are combined to obtain the Atractylodes macrocephala-processed Strychnos nux-vomica decoction for later use.

[0044] Prepare all the decoctions to a concentration of 6 mg / mL. -1 The concentration.

[0045] 3. Animal grouping, model establishment, and drug administration

[0046] Rats were grouped: After one week of acclimatization feeding, the rats' body weight was recorded daily, and the rats were divided into 6 groups of 10 rats each, for a total of 60 rats. These groups were: blank control group (K), model group (M), positive control group [methotrexate (Y)], and raw strychnine group (60 mg / kg). -1 (S), Atractylodes macrocephala and Strychnos nux-vomica group 60mg·kg -1 (L), Atractylodes macrocephala + Strychnos nux-vomica group 60mg·kg -1 (BM).

[0047] Model establishment: After one week of acclimatization, rats in the blank control group were injected subcutaneously with 0.1 mL of physiological saline in the right hind paw, while rats in the other five groups were injected with 0.1 mL of complete Freund's adjuvant (CFA) in the same manner to establish an adjuvant-induced arthritis rat model. After inflammation, varying degrees of swelling appeared in the right paw of the rats, with swollen and red toes and limited movement, indicating successful model establishment. The degree of toe swelling and dietary status of the rats were then closely observed.

[0048] Rats were administered the drug on day 7 after modeling. Rats in the raw Strychnos nux-vomica group, the processed Strychnos nux-vomica group, and the group receiving both Strychnos nux-vomica and Atractylodes macrocephala were given the corresponding decoctions by gavage. Rats in the blank control group and the model group were given an equal volume of 0.5% sodium carboxymethyl cellulose. The total volume of drug administered to each group was 10 mL / kg. -1 (Drug concentration is 6 mg / mL) -1 The drugs were administered simultaneously for 21 consecutive days. The positive control group received methotrexate 0.5 mg / kg. -1 The medication was administered every 3 days. All rats were housed in the same environment with free access to water and food. Their weight was measured daily, and the dosage was adjusted accordingly.

[0049] 4. Arthritis score

[0050] Within 7 days of establishing the model, the general behavior and paw swelling of the model rats were closely observed. Seven days after injection of the modeling drug CFA, the degree of joint swelling in the rats was measured, and the arthritis index scores were calculated based on days 1, 4, 7, 10, 13, 16, 19, and 21 of administration.

[0051] The thickness of the rat's ankle joint and foot was measured with vernier calipers, and a score was given based on the thickness. 0 points represented normal, with no signs of swelling in the toes; 1 point represented redness and swelling of the toes, i.e., slight swelling of the little toe joint; 2 points represented redness and swelling of the toes and foot, with mild swelling and erythema of the toe joints and toes; 3 points represented swelling of the toes down to below the ankle joint, with moderate swelling and erythema below the ankle; 4 points represented complete swelling of the entire toe, including the ankle, foot, and toes, and inability to bend.

[0052] 5. Thymus and spleen index measurement

[0053] After blood collection from each group of rats, the thymus and spleen were removed using sterile instruments. The surface blood was washed away with physiological saline, and residual liquid was blotted dry with filter paper before weighing. The weight was recorded using a 0.01% balance, and the thymus index and spleen index were calculated based on the rat's body weight. The calculation formulas are as follows:

[0054] Thymus index = thymus mass (mg) / body mass (g),

[0055] Spleen index = spleen mass (mg) / body mass (g).

[0056] 6. Serum ELISA detection

[0057] Rats were fasted for at least 12 hours but allowed free access to water before the last administration. One hour after the last administration, rats were anesthetized with sodium pentobarbital, and blood was collected from the abdominal aorta. The system was then set at 4°C and 5000 rpm. -1The serum sample was obtained by centrifugation for 10 min and stored at -80℃. The serum levels of TNF-α, IL-1β, and IL-6 were measured using an enzyme-linked immunosorbent assay (ELISA) according to the kit instructions.

[0058] 7. Biochemical indicator testing

[0059] The levels of liver biochemical indicators ALT and AST, as well as kidney BUN and CRE, in the serum of rats in each group were measured using a fully automated biochemical analyzer.

[0060] 8. Statistical Analysis

[0061] Statistical analysis was performed using GraphPad Prism 5.0. Experimental results are presented as mean ± standard deviation. The t-test was used for comparisons between groups, and a p-value < 0.05 was considered statistically significant.

[0062] II. Experimental Results

[0063] 1. Effects of spleen and thymus index

[0064] After establishing the RA model, the rats experienced weight loss and a significant decrease in food intake. After 21 days of treatment, compared with the control group (K), the spleen and thymus indices of the model group rats were significantly increased (P < 0.01, P < 0.05); compared with the model group (M), the spleen and thymus indices of rats in the raw Strychnos nux-vomica (S), Atractylodes macrocephala-processed Strychnos nux-vomica group (L), and Atractylodes macrocephala + Strychnos nux-vomica group (BM) were all significantly decreased (P < 0.01, P < 0.05). (See attached text). Figure 1 .

[0065] 2. Effects on relevant indicators in rats

[0066] 2.1 Effect of the swelling index on rats

[0067] During the experiment, rats in all groups were active, had formed stools, and normal urine color. The control group had normal food and water intake, while the other groups experienced impaired food and water intake due to arthritis in their hind limbs. No swelling or ulceration was observed on the skin surface of any group of rats, but the model group showed swelling and stiffness from the ankle joint to the toes. After establishing the RA model, rats successively developed redness and swelling in their toes, leading to joint enlargement and difficulty walking. Compared with the model group (M), the group treated with Atractylodes macrocephala and Strychnos nux-vomica (L) showed the best relief of arthritis swelling. Figure 2 .

[0068] The overall arthritis scores of RA rats showed a decreasing trend, indicating that the drug treatment could alleviate the arthritis symptoms of RA rats. Among them, the arthritis scores of rats in the Atractylodes macrocephala-processed Strychnos nux-vomica group (L) were significantly lower than those in the model group (M) (P<0.05). Figure 3 .

[0069] 2.2 Effects on inflammation-related markers in rats

[0070] Inflammatory factors play an important role in RA inflammation and joint damage. Compared with the blank group (K), the serum levels of TNF-α, IL-1β, and IL-6 in the model group (M) rats were significantly increased (P<0.0001). Compared with the model group, the serum levels of TNF-α, IL-1β, and IL-6 in the Atractylodes macrocephala-processed Strychnos nux-vomica group (L) rats were significantly decreased (P<0.0001, P<0.05), and the serum levels of TNF-α, IL-1β, and IL-6 in the raw Strychnos nux-vomica group (S) rats were significantly decreased (P<0.0001, P<0.001, P<0.001), as shown in Table 1.

[0071] Table 1. Concentrations of inflammatory factors in the serum of rats in different groups (unit: pg·mL) -1 )

[0072]

[0073]

[0074] Note: Compared with K, #P<0.05, ##P<0.01, ###P<0.001, ####P<0.0001; compared with M, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

[0075] 2.3 Results of Liver and Kidney Biochemical Indicators Measurement

[0076] Compared with the control group, the levels of ALT, AST, BUN, and CRE in the model group were significantly increased (P<0.05); compared with the model group, the levels of ALT and BUN in the Atractylodes macrocephala-processed Strychnos nux-vomica group were significantly decreased (P<0.05, P<0.001), indicating that Atractylodes macrocephala-processed Strychnos nux-vomica protects against liver and kidney damage in rats with rheumatoid arthritis (see Table 2).

[0077] Table 2. Content of liver and kidney biochemical indicators in different groups of rats

[0078]

[0079] Note: Compared with K, #P<0.05, ##P<0.01, ###P<0.001, ####P<0.0001; compared with M, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

[0080] The above examples illustrate that processing Strychnos nux-vomica with Atractylodes macrocephala ensures therapeutic efficacy while reducing the toxicity of Strychnos nux-vomica, thus improving medication safety. Furthermore, processed Strychnos nux-vomica with Atractylodes macrocephala has shown outstanding efficacy in the treatment of rheumatoid arthritis (RA).

[0081] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. A method for processing Strychnos nux-vomica for intervention in rheumatoid arthritis, characterized in that, This involves decocting raw Strychnos nux-vomica in a decoction of Atractylodes macrocephala, filtering the decoction, and drying the residue to obtain processed Strychnos nux-vomica.

2. The method for processing strychnine according to claim 1, characterized in that, The method for preparing the decoction of Atractylodes macrocephala includes the following steps: Atractylodes macrocephala is soaked in water and then decocted for the first time, and filtered to obtain the first filtrate; water is added to the residue again and decocted for the second time, and filtered to obtain the second filtrate; the two filtrates are combined to obtain the decoction of Atractylodes macrocephala.

3. The method for processing strychnine according to claim 2, characterized in that, During the first decoction, the ratio of Atractylodes macrocephala to water was 1g:20mL; the first decoction was carried out by bringing it to a boil over high heat and then simmering it over low heat for 30 minutes.

4. The method for processing strychnine according to claim 2, characterized in that, During the second decoction, the ratio of the filter residue to water is 1g:10mL; the second decoction is to bring to a boil over high heat, then simmer over low heat for 20 minutes.

5. The method for processing strychnine according to claim 1, characterized in that, The raw nux vomica and the aqueous decoction of Atractylodes macrocephala were mixed at a ratio of 1g:60mL.

6. The method for processing strychnine according to claim 1, characterized in that, The soaking time is 30 minutes.

7. The method for processing strychnine according to claim 1, characterized in that, The decoction process involves bringing the mixture to a boil over high heat, then simmering over low heat for 1 hour.

8. The method for processing strychnine according to claim 1, characterized in that, The drying process involves drying at 60°C for 4 hours.

9. A processed strychnine obtained by the method of processing strychnine according to any one of claims 1-8.

10. The use of the processed Strychnos nux-vomica seed as described in claim 9 in the preparation of a medicament for treating rheumatoid arthritis.

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