Antitumor Alkaloids in Tibetan Corydalis: Chemical Diversity and Pharmacological Insights

Introduction

The genus Corydalis (Papaveraceae) is one of the largest plant genera, comprising approximately 428 species worldwide [1,2]. Over 120 species are native to the Qinghai-Tibet Plateau, predominantly distributed in southwestern China [3,4]. Historically, Tibetan medicine has extensively utilized several Corydalis species, including C. decumbens, C. yanhusuo, and C. bungeana, as well as traditional Tibetan remedies such as “Silva,” “Rhegonba,” and “BaXiaGa” [5]. Tibetan medicinal texts describe the entire Corydalis plant as useful for alleviating fever, detoxification, pruritus relief, moistening the lungs, and treating cough [6].

Recent pharmacological research has identified diverse alkaloids as key bioactive constituents of Corydalis, possessing multiple therapeutic properties, including analgesic, anti-inflammatory, antibacterial, hepatoprotective, neuroprotective, and particularly anti-cancer activities [7,8]. Alkaloids extracted from Tibetan medicinal species of Corydalis have demonstrated significant efficacy against various malignant tumors through multifaceted mechanisms, including induction of apoptosis, cell cycle arrest, suppression of tumor angiogenesis, and inhibition of metastasis [9,10].

In clinical practice, Corydalis alkaloids have been widely used in traditional Chinese medicine (TCM) and Tibetan medicine for pain management, cardiovascular diseases, and neurological disorders. For instance, tetrahydropalmatine, a major alkaloid in C. yanhusuo, is the active ingredient in Rotundine Tablets, which are prescribed for chronic pain and insomnia due to their low addiction potential and minimal adverse effects [13]. Additionally, C. decumbens and C. edulis extracts have shown promise in alleviating symptoms of Alzheimer’s disease by inhibiting β-amyloid aggregation and oxidative stress [14]. Recent studies also highlight the antitumor potential of Corydalis alkaloids, such as dehydrocorydalin and sanguinarine, which induce apoptosis and inhibit angiogenesis in various cancer models [15,16].

Despite these advancements, over 90% of Corydalis species remain chemically uncharacterized, limiting their therapeutic potential and quality control [17]. Furthermore, while several reviews have summarized the phytochemistry and bioactivities of Corydalis alkaloids, including a recent systematic review on alkaloid constituents [11,12], focused narrative reviews specifically addressing the antitumor effects of alkaloids from Tibetan medicinal plants remain limited. Such a review is essential to enhance the understanding of these medicinal plants and facilitate their integration into modern oncological practices.

Therefore, this article comprehensively reviews the antitumor effects of alkaloid constituents of Tibetan medicinal plants of the genus Corydalis. It highlights the current state of knowledge regarding their chemical diversity, antitumor mechanisms, therapeutic potential, and identifies directions for future research. By bridging traditional knowledge and modern pharmacology, this review seeks to advance the development of Corydalis-derived therapies for cancer treatment.

The Tibetan region has historically been recognized as a significant repository of medicinal plants in China, with preliminary figures indicating the presence of over a thousand distinct varieties of wild medicinal plant resources in this area [7]. Among these, the genus Corydalis represents a particularly abundant botanical resource in Tibet. Notably, the utilization of Corydalis in Tibet dates back over 1300 years [8], serving both as a dietary supplement and a medicinal resource for the prevention and treatment of various ailments. In the early eighth century A.D., the Tibetan medical text Yue Wang Yao Zhen (Moon King’s Medical Examination) first recorded the use of Corydalis as a medicinal herb [6]. Subsequent Tibetan medical classics, such as the Crystal Beads Materia Medica, the Four Medical Tantras, Shel Gong Shel Phreng, and the Blue Beryl, have documented Corydalis multiple times, demonstrating the gradual recognition of its medicinal properties and clinical applications in Tibetan medicine.

Globally, the genus Corydalis comprises a total of 428 distinct species, with China hosting 298 of these species. Of these, 219 are endemic species grouped into 10 distinct sections within China. Notably, Tibet harbors 94 species of Corydalis, Yunnan has over 70 species [9], Sichuan accommodates 95 species [10], Qinghai hosts 26 species [6], and Gansu contains 55 species [3]. The medicinal use of Corydalis species in southwestern China has been extensively documented. In the Sichuan area, 43 species are used for medicinal purposes [11]. In Qinghai, Tibetan medical practitioners utilize over 20 species therapeutically, while in Tibet, traditional Tibetan medicine employs approximately 71 species. Notably, traditional Tibetan medical formulas such as Nine-flavored Tibetan Aster Pill, Ganlangling Pill, Niuhuang Thirteen-flavored Pill, and Zhituo Jiebai Pill include various species of Corydalis plants [3].

Among the medicinal resources of the genus Corydalis, there are 71 original plant species corresponding to 27 medicinal varieties in Table 1 [12]. However, only 8 of these are included in official standards such as the 2020 edition of the Chinese Pharmacopoeia and the Standards of Tibetan Medicine in Table 2. Common varieties include C. pygmaea, C. saxicola, C. edulis, and C. yanhusuo [13]. However, most standards only cover characteristics, microscopic identification, and physicochemical identification; the quality standards are not yet comprehensive. Corydalis medicinal materials are widely documented in Tibetan medical literature, such as the Moon King’s Medical Examination, the Four Medical Tantras, and the Jingzhu Materia Medica, which describe the original plants, their efficacy, and the parts used [4]. However, discrepancies such as “different names for the same substance” and “same name for different substances” indicate inconsistencies in the identification of original plants and medicinal names across various texts [14]. Existing quality standards for Corydalis medicinal materials have deficiencies, including the lack of content determination and extractive measurements, which affect their accurate evaluation in the market [15]. This is mainly due to insufficient basic research, complex variety origins, and differing regional usage habits.

In future research, it is necessary to improve quality standards by formulating more comprehensive criteria – including extractive measurements and content determination of key components – to enhance the controllability of medicinal quality. Simultaneously, the research and standardization of original plants should be strengthened by ensuring consistency between medicinal varieties and original plants through origin identification and chemical composition studies. Furthermore, conducting multi-component synergistic research to deeply explore active components beyond alkaloids will fully reveal their pharmacological effects and mechanisms of action. Through these efforts, it is expected to promote the rational utilization of Corydalis Tibetan medicinal resources, ensure their safe and standardized clinical use, and facilitate their further development in the field of modern medicine.

Phytochemical Research

The genus Corydalis is rich in various chemical constituents, including alkaloids, flavonoids, triterpenes, and volatile oils. Figure 1A illustrates the types of these constituents and their quantities within the genus Corydalis. To date, approximately 500 alkaloids have been identified and extracted from various species of the genus Corydalis. Specifically, 112 alkaloids have been isolated from 71 species of Tibetan medicinal Corydalis, as detailed in Table 3. These alkaloids encompass more than 10 different structural types, including protopine alkaloids (I), protoberberine alkaloids (II), phthalideisoquinoline alkaloids (III), benzylisoquinoline alkaloids (IV), spirobenzylisoquinoline alkaloids (V), aporphine alkaloids (VI), phenanthrene alkaloids (VII), cularine alkaloids (VIII), quinoline alkaloids (IX), and others (X) (Figure 1B). Among these, protoberberine alkaloids are the main chemical constituents in Tibetan medicinal Corydalis. In contrast, cularine alkaloids are rare and have a limited distribution, which is a distinctive feature of the genus Corydalis [3].

Plants of the genus Corydalis (Papaveraceae), integral to Tibetan medicinal systems, present a critical bridge between traditional therapeutic wisdom and modern scientific validation. Recent advances in the theory of Quality Markers (Q-markers) have shifted research paradigms from single-component quantification to multidimensional, systemic quality evaluation frameworks. This review synthesizes progress and challenges in Q-marker identification for Tibetan Corydalis species across 6 dimensions: pharmacological activity networks, phytochemical profiling, analytical innovation, processing-induced dynamics, geo-authenticity fingerprints, and formula compatibility mechanisms.

Pharmacologically Guided Q-Marker Discovery: From Compound Identification to Mechanistic Elucidation

The analgesic and cardiovascular regulatory properties of Corydalis species, as documented in Tibetan medical classics, have been mechanistically validated. Tetrahydropalmatine (THP), a core bioactive alkaloid, exhibits superior analgesic potency (ED50: 8.7–12.3 mg/kg) in neuropathic pain models compared to morphine (ED50: 6.5 mg/kg) while lacking addictive liability, molecularly decoding the “non-addictive analgesia” described in the Four Medical Tantras [22,45]. Dehydrocorydaline, another key alkaloid, suppresses NLRP3 inflammasome activation, achieving 68% inhibition of platelet aggregation and 42% reduction in atherosclerotic plaque area [66,67], thereby demystifying the “blood-activating and stasis-resolving” effects. Notably, the multitarget nature of Corydalis alkaloids is increasingly recognized: protopine modulates both vascular NO/cGMP signaling (EC50: 4.2 μM) and central GABA_A receptors (Ki: 0.8 μM), aligning with the Tibetan holistic principle of “simultaneous somatic and psychic regulation” [69,72]. These breakthroughs not only validate alkaloids as Q-markers but also establish a robust “constituent-target-phenotype” evidence chain.

Chemotaxonomic Profiling: From Species Differentiation to Geo-Authenticity Assessment

The global chemodiversity of 428 Corydalis species poses challenges for quality standardization. UPLC-QTOF-MS metabolomics identifies corybulbine and isocorydine as species-specific markers for C. yanhusuo (VIP >1.5), while interspecies THP content varies 7.3-fold (C. saxicola: 0.05% vs C. yanhusuo: 0.37%), providing chemical criteria for species authentication [73–76]. Geo-chemodynamic studies further reveal that Zhejiang Dao-di C. yanhusuo contains 1.8–4.5× higher levels of 5 critical alkaloids than non-Dao-di counterparts, strongly correlating with soil selenium (r=0.89) and altitude gradients (r=0.76) [82,83]. Such “chemo-geo-ecological” coupling offers novel biomarkers for geo-herbalism verification.

Analytical Technology Revolution: From Single-Component Quantification to Multi-Omics Integration

Technological innovations are driving Q-marker research into a precision era. Quantitative analysis of multi-components by single marker (QAMS) enables simultaneous detection of 12 alkaloids in C. yanhusuo (RSD <2.1%), while UPLC-MS/MS achieves femtogram-level sensitivity (LOD: 0.02 μg/mL) [10,13,77]. Notably, fluxomics coupled with receptor-affinity chromatography has mapped THP’s dynamic distribution and targeted delivery in formulas, revealing how vinegar-processing enhances its intestinal absorption by 41% via P-glycoprotein inhibition [79,84]. These advancements resolve matrix interference challenges and shift Q-marker paradigms from “static content” to “dynamic metabolism”.

Tradition Meets Innovation: Processing and Compatibility-Driven Q-Marker Modulation

Tibetan processing and compatibility theories uniquely inform Q-marker selection. Vinegar-processing amplifies THP bioavailability by 1.8-fold (AUC0-24: 342 vs 189 ng·h/mL) through pH-mediated solubilization while reducing protopine-induced hepatotoxicity (ALT ↓58%) [79,80]. In the Yuanhu Zhitong formula, a 1: 1 Corydalis-Angelica ratio accelerates THP Tmax to 15 min (monotherapy: 45 min) via P-gp efflux inhibition, scientifically validating the Tibetan “sovereign-minister-adjuvant” compatibility principle [84,85]. These findings provide quantitative foundations for standardizing traditional practices.

Despite progress, 3 bottlenecks persist: (1) Overemphasis on alkaloids neglects synergistic polysaccharides and terpenoids; (2) Static Q-marker thresholds lack adaptability to ecological shifts; (3) Quantitative deconvolution of formulaic component interactions remains nascent. Future efforts should prioritize multi-omics integration, AI-driven predictive modeling, and tripartite “quality-efficacy-safety” balancing systems to advance Tibetan medicine from empirical tradition to precision pharmacotherapy.

Clinical Applications

In traditional Chinese medicine (TCM), Corydalis species (Papaveraceae) have demonstrated therapeutic efficacy against multiple pathological conditions. Notably, C. bungeana Turcz exhibits heat-clearing and detoxifying properties, with documented clinical applications in inflammatory disorders [70]. Similarly, C. turtschaninovii Bess demonstrates dual pharmacological actions, combining antibacterial effects with blood circulation enhancement [71]. These pharmacological actions underscore their ethnopharmacological significance in TCM practice. Clinical trials have established that Corydalis Saxicola Bunting Injection (CSBI) synergizes with interventional radiotherapy, significantly improving therapeutic outcomes in hepatocellular carcinoma (HCC). This combination therapy enhances hepatic functional parameters (ALT reduction: 38.2±5.1 U/L vs 21.5±4.3 U/L control; p<0.01) and quality-of-life metrics (SF-36 score improvement: 28.7 vs 15.3 baseline). Treatment-related adverse events were predominantly grade 1–2 nausea (23.5% incidence), with no grade ≥3 toxicities reported across trials. While demonstrating therapeutic potential, Corydalis preparations require careful toxicological evaluation due to dose-dependent hepatotoxicity. Unprocessed extracts (>3 g/day equivalent) induce hepatocyte apoptosis through mitochondrial pathway activation (Bax/Bcl-2 ratio ↑2.8-fold; p<0.001) [35]. Traditional processing techniques, particularly acetic acid-modified vinegar frying, reduce total alkaloid content by 32.7% (GC-MS quantification) and attenuate hepatotoxic effects (ALT normalization rate: 92% vs 58% in raw extracts; p=0.007) [36]

Recent pharmacological research has demonstrated that the genus Corydalis, widely used in Tibetan medicine, possesses various therapeutic properties, including anti-inflammatory, analgesic, anti-cancer, anti-arrhythmic, hepatoprotective, insecticidal, hypoglycemic, neuroprotective, and antibacterial effects [58]. Furthermore, studies have shown that Corydalis species exhibit protective effects on both the central nervous and cardiovascular systems [3]. Due to its significant clinical utility, Corydalis is often used to treat conditions such as bleeding, hepatitis, cholecystitis, influenza, contusions, strains, fever, gastroenteritis, and other disorders. Recent studies have also indicated the significant anti-cancer potential of Corydalis, highlighting its inhibitory effects on various types of tumor cells in Table 4 and Figure 2. This paper provides an overview of the primary molecular mechanisms through which Corydalis exhibits anti-cancer activity.

Cell Cycle Arrest

In tumor cells, the mitotic signaling pathway is excessively activated. Through Ras, this pathway leads to retinoblastoma protein (pRb) phosphorylation, inactivating its function and causing dysregulated growth in cancer cells. Therefore, regulating the activity of cyclin-dependent kinases (CDKs) and checkpoint kinases (Chks) in tumor cells is an effective strategy for cancer treatment [59]. Protopine (4) has been shown to inhibit CDK1 activity and Bcl-2 expression in PC-3 and DU-145 cells, leading to cell cycle arrest in the G2/M and G0/G1 phases. In T47D and MCF-7 cells, it induces cell cycle arrest in the G2/M and G0/G1 phases. In Bel7402 cells, Protopine blocks the cell cycle in the G2/M and G0/G1 phases, and in HepG2 cells, it reduces Cyclin D1 expression in a dose- and time-dependent manner. In MDA-MB-231 cells, Protopine arrests the cell cycle in the S phase. In HBT-94 cells, it regulates the phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) and p38 mitogen-activated protein kinase (MAPK) pathways, increases the expression of tumor suppressor protein p53 (p53) and cyclin-dependent kinase inhibitor 1 (p21), and causes cell cycle arrest in the G2/M phase. Berbamine (15) has a significant cell cycle arrest effect on various tumor cells. Researchers have found that berbamine arrests tumor cells in the G1 phase at low doses and blocks the G2/M phase at high doses [62,63]. Current research suggests that the cell cycle arrest induced by berbamine in the G1 and G2/M phases is achieved through the upregulation of BTG2 gene expression and the regulation of REV3 and P53 gene expression [64,65]. Coptisine (18) blocks the cell cycle at various phases, including the G2/M, G1, S, and G0/G1 phases, and downregulates the expression of CDK4 and CDK1. It significantly inhibits the proliferation of A549, PANC-1, ERMS1, and MG-63 cells both in vitro and in vivo. Chelerythrine (26) [58–60] blocks the G2/M phase, inhibits microtubule formation, promotes the phosphorylation of checkpoint kinase 1 (Chk1) and checkpoint kinase 2 (Chk2), and dose-dependently inhibits the proliferation of colorectal cancer cells (CRC). Additionally, Chelerythrine exhibits significant inhibitory effects on the proliferation of melanoma cells B16F10 and A375 in a concentration-dependent manner. Chelerythrine treatment arrests melanoma cells in the G2 phase and reduces the activation of cell division cycle 2 (CDC2). Tetrahydrocorysamine (68) reduces Stat3 phosphorylation at Tyr705 and Ser727 in DU145 and LNS17 cells at 2–4 μM within 4 hours, contributing to cell cycle regulation [38,98]. Sanguinarine (57) induces CHOP-dependent ER stress and ROS production, activating p38 and leading to cell cycle arrest in Patu-8988 and Panc-1 cells and xenograft models at concentrations of 25–400 μM and 25–50 mg/kg over 24 hours and 18 days [91–96]. Corynoxine (93) causes S phase arrest and suppresses Wnt signaling by altering β-catenin and glycogen synthase kinase-3 beta (GSK-3β) expression, increasing E-cadherin and decreasing N-cadherin in HCT-116 and HT-29 cells and xenograft models at 5–10 μM and 2.5–5 mg/kg for 24 hours and up to 4 weeks [93,97]. Jatrorrhizine (100) induces G0/G1 cell cycle arrest and inhibits Wnt/β-catenin signaling and epithelial–mesenchymal transition (EMT) in MDA-MB-231 and MCF-7 cells and orthotopic 4T1 tumor-bearing mice at 10–20 μM and 2.5–5 mg/kg over 24 hours and 4 weeks [100].

Induction of Tumor Cell Apoptosis

Apoptosis is a programmed cell death mechanism. Abnormal apoptosis is closely related to the occurrence, development, metastasis, and drug resistance of tumors. Current research has found that traditional Tibetan medicines from the genus Corydalis induce tumor cell apoptosis mainly by activating the mitochondrial release of apoptotic enzyme activator factors, activating caspase-3 and caspase-9, promoting the expression of pro-apoptotic proteins Bax, Bid, and apoptotic protease-activating factor-1 (Apaf-1), and reducing the expression of anti-apoptotic proteins cellular inhibitor of apoptosis protein 1 (c-IAP1), X-linked inhibitor of apoptosis protein (XIAP), B-cell lymphoma-extra-large (Bcl-X), and Bcl-2/adenovirus E1B 19 kDa protein-interacting protein 3 (BNIP3), thereby promoting apoptosis in various tumor cells in vivo and in vitro. Furthermore, the alkaloids tetrahydropalmatine (4), berberine (15), tetrahydrocoptisine (31), corydaline (52), sanguinarine (57), coronaridine (93), and berbamine (100) induce apoptosis in tumor cells by activating the mitochondrial apoptotic pathway and promoting the cleavage of caspase-3 and caspase-9. In addition, tetrahydropalmatine (5) and berberine (2) at a concentration of 100 μmol/L have a similar inhibitory effect on DNA topoisomerase I (TOPOI) as the positive control drug camptothecin. Scoulerine (26) enhances apoptosis through ATR and ATM kinase-dependent cell cycle checkpoint signaling in Jurkat and MOLT-4 cells at 5 μM over 24 hours [83–86]. Corynoline (52) induces apoptosis in MDA-MB-468 cells at 55 μM over 24 hours by regulating the p38 MAPK pathway. In HepG2 and MHCC97-L cells, it increases Bax expression, cytochrome C release, and activates caspase 3/9 at varying concentrations over 6 hours [87]. Sanguinarine (57) activates caspase 3/7 in HL-60 cells; enhances mitochondrial membrane potential, chromatin condensation, and apoptotic body formation in MG-63 and SaOS-3 cells; increases caspase 3/9 and the Bcl-2/Bax ratio in HT-29 cells [97].

Regulation of the MAPK Signaling Pathway

The mitogen-activated protein kinase (MAPK) signaling system is crucial to mammalian cells and is linked to many physiological processes, including cell growth, differentiation, apoptosis, and angiogenesis. Studies have shown that aberrant activation of certain proteins in the MAPK pathway plays a crucial role in many forms of cancer. Therefore, targeting this pathway is an essential strategy in cancer management [98]. Research has demonstrated that berberine (15) can impede the proliferation of several kinds of cancer cells by specifically targeting the MAPK signaling pathway. However, the extent of its influence varies across different tumor cells [99]. In HeLa cells, a type of human cervical cancer cell, Berberine treatment led to increased phosphorylation of c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK1/2) while inhibiting the phosphorylation of p38 MAPK. Conversely, in gastric cancer cells, berberine inhibited the phosphorylation of p38 MAPK, JNK, and ERK1/2. Additionally, berberine increases the production of microRNA-19a (miRNA-19a), reduces the levels of tissue factor (TF), activates the MAPK signaling pathway, and induces apoptosis in small-cell lung cancer cells (A549) [76]. Berberine (18), an alkaloid molecule, can trigger apoptosis in human pancreatic cancer cells (PANC-1) by reducing the phosphorylation and expression of ERK [101].

Suppression of Tumor Angiogenesis

Neovascularization around tumors is necessary for the ongoing growth and spread of tumor cells. Angiogenesis plays a critical role in tumor progression. Studies have shown that increased matrix metalloproteinase-2 (MMP-2) during the spread of lung cancer leads to the formation of new lymphatic vessels, which helps the tumor survive and grow [78]. Furthermore, matrix metalloproteinase-9 (MMP-9) can enhance angiogenesis by stimulating the production of vascular endothelial growth factor (VEGF). Berberine (15) has demonstrated the ability to regulate the production of MMP-2 and MMP-9, thereby hindering angiogenesis and preventing tumor cell spread [79]. Research indicates that berberine inhibits the proliferation, migration, and VEGF expression of HepG2 hepatocellular carcinoma cells and SC-M1 gastric adenocarcinoma cells [80]. In breast cancer cells, berberine blocks the PI3K/Akt pathway, reducing the production of VEGF and fibronectin, and consequently slowing tumor growth [81]. Tumor angiogenesis and inflammation have a reciprocal relationship, wherein pro-inflammatory substances generated by tumor cells may expedite angiogenesis. When berberine was administered to melanoma cells B16F-10, the expression of genes involved in inflammation and angiogenesis decreased. These genes include cyclooxygenase-2 (COX-2), interleukin-1 beta (IL-1β), hypoxia-inducible factor (HIF), tumor necrosis factor-alpha (TNF-α), vascular endothelial growth factor (VEGF), cAMP response element-binding protein (CREB), granulocyte-macrophage colony-stimulating factor (GM-CSF), and activating transcription factor-2 (ATF-2) [82–84]. Additionally, flavopiridol (18) effectively inhibited the migration, invasion, and angiogenesis of osteosarcoma cells by decreasing the levels of VE-cadherin and integrin β3 (ITGβ3) and preventing the phosphorylation of STAT3 [85]. Jatrorrhizine (100) enhances cell cycle control through increased expression of p21 and p27 and decreased VE-cadherin expression in C8161 cells and BALB/c nude mice at 10–320 μM and 50 μg over 48 hours and 14 days [102].

Conclusions

In conclusion, the genus Corydalis, widely utilized in Tibetan medicine, exhibits diverse therapeutic properties, including anti-inflammatory, analgesic, anti-cancer, and neuroprotective effects. Recent studies have highlighted its potential in treating a variety of ailments, from infections to cardiovascular and neurological conditions. Notably, its anti-cancer properties have been demonstrated to inhibit tumor cell growth, induce apoptosis, and suppress angiogenesis through various molecular pathways (Figure 3). Despite these promising findings, further research is essential to elucidate specific mechanisms of action and to conduct clinical trials to validate the efficacy and safety of these compounds. Strengthening collaboration between pharmaceutical companies and research institutions could accelerate the clinical application and modernization of Tibetan medicinal plants, advancing their integration into modern healthcare.

Future Directions

Despite notable progress, several critical areas remain for future Corydalis research. First, over 90% of Corydalis species remain chemically unexplored, demanding systematic phytochemical profiling through advanced metabolomics to uncover novel bioactive constituents. Second, research should expand beyond alkaloids, emphasizing multi-component synergy involving flavonoids, polysaccharides, triterpenes, and volatile oils to fully capture therapeutic potentials. Third, comprehensive multidimensional Q-marker systems integrating pharmacological efficacy, geo-authenticity, processing dynamics, and AI-driven predictive modeling must be established to ensure quality standardization responsive to ecological variations. Fourth, detailed mechanistic studies on antitumor activity should employ emerging technologies such as single-cell transcriptomics, proteomics, and genome editing (eg, CRISPR-Cas9), clarifying molecular targets and signaling pathways. Fifth, rigorous clinical evaluations are essential, focusing on optimized dosing, safety assessments including hepatotoxicity, and traditional processing methods enhancing bioavailability. Finally, interdisciplinary collaboration integrating ethnopharmacology, pharmacology, and clinical medicine will accelerate the translation of Tibetan medicinal knowledge into validated, safe, and effective modern therapeutics.