Application of exocarpium citri grandis extract in preparation of anti-pulmonary nodule product
By treating Citrus reticulata extract with an aqueous solvent, an anti-pulmonary nodule drug was prepared, which solved the shortcomings of existing pulmonary nodule treatment technologies. It achieved significant improvement in the inhibition of MWCNT-induced pulmonary nodules and inflammatory infiltration, providing a gentler treatment option and expanding the application field of Citrus reticulata.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- JIANMA PHARM (GUANGDONG) CO LTD
- Filing Date
- 2026-04-10
- Publication Date
- 2026-05-19
AI Technical Summary
There is a lack of effective and gentle treatment methods for pulmonary nodules in the current technology, especially for early-stage pulmonary nodules. Furthermore, traditional Chinese medicine lacks a unified syndrome standard in this regard, which may lead to missing the best treatment time or causing unnecessary waste of medical resources.
Citrus reticulata extract, especially its aqueous extract, was prepared into liquid or solid dosage forms by heating and reflux extraction and concentration. These were used to prepare anti-pulmonary nodule drugs and significantly inhibited the development of MWCNTs-induced pulmonary nodules in mouse models.
Citrus reticulata extract can significantly inhibit MWCNT-induced pulmonary nodule formation and inflammatory infiltration, reduce the area of nodular lesions, provide a gentler treatment for pulmonary nodules, broaden the application scope of Citrus reticulata, and has the potential to prevent and treat lung cancer.
Smart Images
Figure CN122056953A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to novel uses of Citrus reticulata extract, and more specifically, to the use of Citrus reticulata extract in the preparation of anti-pulmonary nodule products. Background Technology
[0002] Pulmonary nodules are focal, round or oval-shaped, solid or subsolid lung shadows with increased density, appearing on imaging as a diameter of 3 cm or less. In recent years, with the development of imaging technology and increased emphasis on physical examinations, the detection rate of pulmonary nodules has significantly increased. A 2021 analysis of pulmonary nodule detection rates showed that 4,817,014 pulmonary nodules were detected among 10,202,034 individuals undergoing physical examinations, with a detection rate of 47.2% (standardized detection rate of 40.6%). The detection rate was 47.4% (standardized detection rate of 40.4%) in men and 47.0% (standardized detection rate of 40.7%) in women. A study analyzed data from 9,167,999 individuals undergoing physical examinations across 31 provinces, autonomous regions, and municipalities in China (excluding Hong Kong, Macau, and Taiwan). The results showed a lung nodule detection rate of 55.9% (standardized detection rate of 54.3%). Overall, in recent years, depending on the scope and subjects of the surveys, the detection rate of lung nodules has ranged from 20% to 60%, indicating a relatively high detection rate.
[0003] Pulmonary nodules have an insidious onset, accompanied by persistent chronic inflammation, and can gradually develop into pulmonary fibrosis, or even lung cancer leading to death. Approximately 95% of pulmonary nodules are benign. Inflammation caused by fungal or bacterial infections, or pulmonary vascular abnormalities, can lead to the formation of solitary pulmonary nodules. Long-term exposure to coal dust or mineral dust (silica or beryllium), or blood clots mixed with bacteria entering the lungs through the bloodstream, can cause multiple pulmonary nodules. However, the formation mechanism of pulmonary nodules is not yet fully understood.
[0004] Currently, Western medicine lacks specific treatments for early-stage pulmonary nodules. It primarily relies on regular follow-up chest CT scans to monitor small nodules or those with uncertain morphology, which carries the risk of missing the optimal treatment window. For solid nodules with a diameter of 15 mm or greater, or those exhibiting malignant CT signs (such as lobulation, spiculation, pleural traction, air-filled bronchioles, cavitation, and eccentric thick-walled cavities) and a diameter between 8 mm and 15 mm, as well as mixed pulmonary nodules larger than 8 mm, the main treatment methods include surgical resection, stereotactic radiosurgery, and thermal ablation. This inevitably leads to the overuse of pulmonary nodule surgery. For example, postoperative pathological examinations in some patients reveal benign lesions, which not only harms the patient but also results in unnecessary waste of medical resources. Traditional Chinese medicine (TCM) clinical diagnosis and treatment largely rely on the clinical experience of individual physicians, lacking unified TCM syndrome differentiation standards for the diagnosis and treatment of pulmonary nodules. TCM remains in the exploratory stage in the diagnosis and treatment of pulmonary nodules; therefore, developing more effective and gentler treatment methods for pulmonary nodules is of great significance.
[0005] Chinese herbal medicine Huajuhong ( Exocarpium Citri Grandis Citrus reticulata peel (ECG) is the dried outer peel of the immature or nearly mature fruit of the pomelo or citrus fruit plant (Citrus reticulata var. huazhouensis), also known as "hairy tangerine peel," "yellow seven-claw peel," "glossy green seven-claw peel," or "seven-claw red peel." Citrus reticulata peel prefers a warm and humid climate, is intolerant of drought, and has poor cold resistance, requiring an average annual temperature above 16℃. Therefore, it is mainly produced in Huazhou City, Guangdong Province. Many medical books record that Citrus reticulata peel has the functions of resolving phlegm, relieving cough, regulating qi, and soothing the stomach. For example, the Qing Dynasty's *Compendium of Materia Medica Supplement* records: "Citrus reticulata peel is miraculously effective in treating phlegm, eliminating greasy food accumulation, and relieving hangovers and soothing the stomach..." During the Ming and Qing dynasties, Huajuhong (a type of tangerine peel) enjoyed a reputation as a "rare and precious medicine for treating coughs and protecting the lungs," and was listed as a tribute to the imperial court. In modern clinical practice, it is often used to treat coughs with excessive phlegm, food stagnation and alcohol poisoning, nausea and vomiting, chronic bronchitis, vomiting and diarrhea, asthma, and coughs caused by wind-cold and colds with itchy throat and excessive phlegm. It has various physiological effects such as anti-inflammatory and anti-tumor effects. However, there are no reports of Huajuhong being used to prepare drugs for treating pulmonary nodules. Therefore, Huajuhong and its active ingredients show great potential in the preparation of drugs for treating pulmonary nodules. Summary of the Invention
[0006] The technical problem to be solved by the present invention is to overcome the defects and deficiencies of the existing technologies and to provide the application of Citrus reticulata extract in the preparation of anti-pulmonary nodule products.
[0007] Therefore, the purpose of this invention is to provide the application of Citrus reticulata extract in the preparation of anti-pulmonary nodule drugs.
[0008] Preferably, the Citrus reticulata extract is a polar solvent extract of Citrus reticulata, and more preferably, it is an aqueous extract of Citrus reticulata.
[0009] Preferably, the aqueous extract of Citrus reticulata is obtained by reflux extraction of Citrus reticulata medicinal material with water, filtration and combination of filtrates, and concentration.
[0010] Further optimization involves adding the Citrus reticulata extract to water at a mass ratio of 1:10, heating and refluxing for 2-3 times, each time for 1-2 hours, filtering and combining the filtrates, and then concentrating the extract.
[0011] Preferably, the animal model of the lung nodule is constructed using MWCNTs.
[0012] Preferably, the drug can be formulated into a liquid dosage form or a solid dosage form. The liquid dosage form is a solution, suspension, or emulsion, and the solid dosage form is a tablet, capsule, pill, powder for injection, sustained-release preparation, etc.
[0013] The present invention has the following beneficial effects:
[0014] (1) This invention is the first to discover that the extract of Citrus reticulata has the effect of alleviating pulmonary nodular lesions induced by MWCNTs and inhibiting the development of pulmonary nodules.
[0015] (2) Specifically, this invention administered a certain concentration of Citrus reticulata extract to mice with MWCNT-induced pulmonary nodules via gavage. It was found that the aqueous extract of Citrus reticulata significantly inhibited MWCNT aggregation and the resulting inflammatory infiltration, while also reducing the area of MWCNT-induced nodular lesions and inhibiting the formation of pulmonary nodules. Citrus reticulata is expected to become a milder drug for inhibiting the development of pulmonary nodules.
[0016] Therefore, this invention is the first to discover, through research, that simply inhibiting inflammation (the effect of dexamethasone) cannot effectively improve the imaging characteristics (CT values) of pulmonary nodules, while the aqueous extract of Citrus reticulata peel showed a significant nodule-improving effect. This suggests that Citrus reticulata peel may inhibit the development of pulmonary nodules by acting on MWCNTs-induced pulmonary nodules through multiple targets, and holds promise for the preparation of drugs to combat the occurrence and development of pulmonary nodules. Compared with existing technologies, the preparation method of Citrus reticulata peel extract in this invention is simple and low-cost, and as a natural plant extract, it can be used as a preventive health food or therapeutic drug. This invention not only provides a new direction for the treatment of pulmonary nodules, but also broadens the application scope of Citrus reticulata peel, revealing its broad application prospects in maintaining human health and preventing lung cancer. Attached Figure Description
[0017] Figure 1 The results of micro-CT scans of the lungs in each group of animal experiments are shown. A: CT images; B: Statistical analysis of CT values (mean ± standard deviation, n=6).
[0018] Figure 2 HE staining results of lung tissue sections from each group in animal experiments are shown. Detailed Implementation
[0019] The present invention will be further described below with reference to specific embodiments, but the embodiments do not limit the invention in any way. Unless otherwise specified, the reagents, methods and equipment used in the present invention are commonly used in the field of repair.
[0020] Unless otherwise specified, all reagents and materials used in the following examples are commercially available.
[0021] The Huajuhong (a type of tangerine peel) used in this embodiment of the invention is produced in Nawu Town, Huazhou City, Guangdong Province.
[0022] The C57 BL / 6J male mice used in this invention were purchased from Zhuhai Beston Biotechnology Co., Ltd. and raised at the Animal Research Platform of the School of Life Sciences, Sun Yat-sen University. The experimental animal use license number is SYXK (Guangdong) 2025-0313, and the ethics number is SYSU-LS-IACUC-2025-0009.
[0023] Example 1: Preparation of aqueous extract of Citrus reticulata peel
[0024] Weigh 35.1 g of Citrus reticulata extract and place it in a round-bottom flask. Add 10 times the amount of water according to the mass ratio of Citrus reticulata extract and heat under reflux for 3 times, 1 hour each time. Filter, combine the filtrates and perform vacuum filtration. Concentrate the filtered liquid to prepare a concentrated solution with a concentration of 702 mg / mL (based on the amount of raw drug). This concentration is determined by converting the human dosage of 6 g / d in the Chinese Pharmacopoeia to 9 times the mouse dosage.
[0025] Example 2: Study on the anti-pulmonary nodule effect of Citrus reticulata extract
[0026] The following experiments were conducted using the water extract of Citrus reticulata obtained in Example 1.
[0027] 1. Experimental Grouping: After 7 days of acclimatization, the purchased male C57 BL / 6J mice were randomly divided into 4 groups of 6 mice each: control group, model group, dexamethasone group, and Citrus reticulata peel group. During the experiment, all mice had free access to food and water, and their weight was measured every other day. Specific grouping procedures are as follows:
[0028] (1) Control group: The same operation as the model group, but the same volume of 35% surfactant / saline without multi-walled carbon nanotubes (MWCNTs) was dripped into the model group. After dripping, physiological saline was administered by gavage at a volume of 0.1 mL / 10 g for 30 days.
[0029] (2) Model group: After sedating mice with isoflurane, the airway was instilled and the mice were fixed on the tracheal intubation platform. A pen-type spotlight was placed against the mouse's throat to open the mouse's mouth. The mouse's tongue was pulled out as much as possible and the epiglottis was located. The mouse's epiglottis was intubated on the glottic side using an indwelling intravenous needle. An insulin injection needle with a capillary tube was connected through the indwelling intravenous needle. 50 μl of 35% surfactant / saline solution containing 100 μg MWCNT was instilled to construct a lung nodule model. 30 days after the instillation, physiological saline was administered by gavage at a dose of 0.1 mL / 10 g for 30 days.
[0030] (3) Dexamethasone group: The same procedure as the model group was performed. A lung nodule model was constructed by dripping 50 μl of a 35% surfactant / saline solution containing 100 μg MWCNT. After dripping, dexamethasone (concentration of 0.1 mg / mL) was administered by gavage at a dose of 0.1 mL / 10 g for 30 days.
[0031] (4) Huajuhong group: The same operation as the model group was performed. 50 μl of 35% surfactant / saline solution containing 100 μg MWCNT was dripped into the lung nodule model. After dripping, Huajuhong water extract (concentration of 702 mg / mL based on crude drug amount) was administered by gavage at a volume of 0.1 mL / 10 g for 30 days.
[0032] 2. Experimental Methods and Results
[0033] 2.1 Observation of lung micro-CT imaging
[0034] Lung CT, due to its extremely high spatial and density resolution, has become the gold standard for clinical detection, diagnosis, and follow-up of pulmonary nodules. To simulate this clinical scenario in an animal model and achieve in vivo, longitudinal, and quantitative assessment of lesion growth and treatment response, this study used high-resolution micro-CT scanning as the core imaging monitoring method. Different tissue types have different CT values. Generally, the CT value of water is defined as 0 HU, and the CT value of air is defined as -1000 HU. Because normally inflated lung tissue contains a large amount of air, the CT value is low, usually negative. If nodular lesions occur in the lungs, the CT value will increase, even becoming positive. This was used to determine whether nodules had formed in the model group and to compare the differences between groups. Specific scan results (30 days after drug administration) are as follows: Figure 1 The results showed that the lungs of the model group developed lesions with ground-glass opacities. After administration of the anti-inflammatory drug dexamethasone, the nodule area did not change significantly, and the CT value decreased but there was no statistical difference. However, after administration of Citrus reticulata extract, the area of the opacity decreased significantly, and the CT value was significantly lower than that of the model group, indicating that Citrus reticulata extract has the effect of inhibiting the development of lung nodules.
[0035] 2.2 HE staining observation of tissue sections
[0036] Mice were sacrificed after 60 days. Lung tissue was fixed in 4% paraformaldehyde, embedded in paraffin, and cut into 4 μm sections. After staining with hematoxylin and eosin, the pathological condition of the lung tissue was observed under a microscope, and differences between groups were compared. Specific results are shown below. Figure 2 As shown in the figure, the results indicated that MWCNTs aggregated in the lung tissues of the model group, dexamethasone group, and Citrus reticulata group. The model group exhibited typical nodular lesions. After dexamethasone administration, inflammatory cell infiltration in the lung tissue was reduced, and the degree of MWCNT aggregation and the area of nodular lesions decreased slightly, but the improvement was limited. In contrast, the Citrus reticulata group significantly inhibited inflammatory infiltration in the lungs, significantly reduced MWCNT aggregation in the lung tissue, and shrunk the area of nodular lesions.
Claims
1. Application of Citrus reticulata extract in the preparation of anti-pulmonary nodule drugs.
2. The application according to claim 1, characterized in that, The aforementioned Citrus reticulata extract is a polar solvent extract of Citrus reticulata.
3. The application according to claim 2, characterized in that, The Citrus reticulata extract is an aqueous extract of Citrus reticulata.
4. The application according to claim 3, characterized in that, The aqueous extract of Citrus reticulata is obtained by reflux extraction of Citrus reticulata extract with water, filtration and combination of filtrates, and concentration.
5. The application according to claim 4, characterized in that, The aqueous extract of Citrus reticulata is obtained by adding Citrus reticulata medicinal material to water at a mass ratio of 1:10, heating and refluxing for 2-3 times, each time for 1-2 hours, filtering and combining the filtrates, and then concentrating.
6. The application according to claim 1, characterized in that, The lung nodules described were constructed using MWCNTs as an animal model.
7. The application according to claim 1, characterized in that, The drug is formulated into a liquid dosage form or a solid dosage form.
8. The application according to claim 7, characterized in that, The liquid dosage form is a solution, suspension, or emulsion.
9. The application according to claim 7, characterized in that, The solid dosage form is a tablet, capsule, pill, powder for injection, or sustained-release preparation.