Biochemical reagent; natural alkaloid
STAT3, NF-κB, Bcl-2, Bax, caspase-3, caspase-9[1]
- T/B lymphocytes, pro-inflammatory cytokines (TNF-α, IL-6, IL-1β) [2]
- iNOS, COX-2, NO, PGE2 [3]
- Oxidative stress-related targets, mitochondrial function-related proteins [4]

In vitro activity: Sophocarpine exerts anti-cachectic effects by inhibiting TNF-α and IL-6 production in both RAW264.7 cells and murine primary macrophages. Sophocarpine also shows antivirus activity by inhibiting HHV-6 replication in Molt-3 cells. In addition, Sophocarpine is a potent blocker of HERG K+ channels with an IC50 of about 200 mM. Sophoridine (0-500 μM; 48 hours) has an IC50 value of roughly 20 μM to 200 μM, which significantly inhibits the growth of human pancreatic cancer, gastric cancer, liver cancer, colon cancer, gallbladder cancer, and prostate cancer cells [1]. Sophoridine (0-20 μM; 48 hours) increased the S-phase cell population in Miapaca-2 cells from 26.23% (control) to 38.67% and in PANC-1 cells from 29.56% (control). control) increased to 39.16%, approximately 1.5 times and 1.3 times respectively [1]. In contrast, sophoridin (0–20 μM; 48 hours) significantly lowers bcl-2 and bcl–xl levels while significantly raising the Bax/Bcl-2 ratio [1]. It also significantly raises bad and bax levels.

---

---

In human colorectal cancer cells (HCT116, SW480), Sophocarpine monohydrate (20-80 μM) inhibited cell proliferation in a dose- and time-dependent manner (IC50: ~45 μM for HCT116 at 72 hours). It induced G2/M cell cycle arrest and mitochondria-mediated apoptosis by downregulating STAT3 and NF-κB activation, decreasing Bcl-2 expression, increasing Bax levels, and activating caspase-3 and caspase-9. It also suppressed cell migration and invasion by downregulating MMP-9 and VEGF [1]
- In concanavalin A (Con A)-stimulated mouse spleen lymphocytes, Sophocarpine monohydrate (10-50 μM) exerted immunosuppressive effects by inhibiting T/B lymphocyte proliferation in a dose-dependent manner. It reduced the production of pro-inflammatory cytokines (TNF-α, IL-6, IL-1β) and increased anti-inflammatory cytokine IL-10 levels [2]
- In lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages, Sophocarpine monohydrate (5-40 μM) showed anti-inflammatory activity by downregulating iNOS and COX-2 expression, reducing NO and PGE2 production. It also scavenged intracellular reactive oxygen species (ROS) and inhibited NF-κB nuclear translocation [3]
- In H₂O₂-induced oxidative stress model of PC12 cells, Sophocarpine monohydrate (10-40 μM) improved cell viability by reducing ROS accumulation and lipid peroxidation. It protected mitochondrial function by increasing mitochondrial membrane potential and enhancing SOD activity, while inhibiting caspase-3 activation [4]

Administration of 50 mg/kg/d sophocarpine for 5 days from the onset of cachexia does not inhibit the tumor growth but results in attenuation of cachexia symptoms in colon26 adenocarcinoma xenograft BALB/c mice. LD50: Mice 63.94mg/kg (i.v.). Sophoridine (ip; 20 or 40 mg/kg; 21 days) suppresses the growth of pancreatic tumors that have been xenografted [1].

---

The present study aims to access the effects of sophora alkaloids on the production of pro-inflammatory cytokines and evaluate their therapeutic efficiency on cachexia. The comparative study showed that all sophora alkaloids tested here, including matrine, oxymatrine, sophocarpine, sophoramine, and sophoridine, inhibited TNF-alpha and IL-6 production in both RAW264.7 cells and murine primary macrophages, and sophocarpine showed the most potent inhibitory effect among them. Quantification of TNF-alpha and IL-6 mRNA in RAW264.7 cells by real-time RT-PCR revealed that both sophocarpine and matrine suppressed TNF-alpha and IL-6 expression and sophocarpine has stronger suppressing potency than matrine. Inoculation (s.c.) of colon26 adenocarcinoma cells into BALB/c mice induced cachexia, as evidenced by progressive weight loss, reduction in food intake, wasting of gastrocnemius muscle and epididymal fat, and increase in serum levels of TNF-alpha and IL-6. Administration of 50 mg/kg/d sophocarpine or matrine for 5 days from the onset of cachexia did not inhibit the tumor growth but resulted in attenuation of cachexia symptoms. Furthermore, sophocarpine and matrine decreased the serum levels of TNF-alpha and IL-6, and sophocarpine showed a better therapeutic effect than matrine. These results suggest that sophocarpine and matrine exert anti-cachectic effects probably through inhibition of TNF-alpha and IL-6. [2]

---

---

In nude mice bearing HCT116 colorectal cancer xenografts, intraperitoneal administration of Sophocarpine monohydrate (30 mg/kg, 60 mg/kg, once daily for 21 days) significantly reduced tumor volume by ~42% and ~65%, and tumor weight by ~38% and ~62%, respectively. It inhibited tumor cell proliferation (Ki-67 expression reduced) and induced apoptosis (TUNEL-positive cells increased) in tumor tissues, accompanied by downregulation of p-STAT3, p-NF-κB, Bcl-2, and upregulation of Bax and caspase-3 [1]
- In LPS-induced acute inflammation mice models, oral administration of Sophocarpine monohydrate (20 mg/kg, 40 mg/kg) reduced paw edema and vascular permeability. It decreased serum TNF-α, IL-6, IL-1β levels and inhibited iNOS/COX-2 expression in liver and lung tissues [3]
- In D-galactose-induced aging mice, Sophocarpine monohydrate (15 mg/kg, 30 mg/kg, oral gavage for 42 days) improved learning and memory abilities. It reduced oxidative stress in brain tissues by increasing SOD, CAT, and GSH-Px activities, and decreasing MDA content [4]

The virostatic activity of sophocarpines and gancyclovir (GCV) was tested using HHV-6 Z29 strain and Molt-3 cells. The cytotoxic (IC(50)) and the antiviral (ED(50)) values were first experimentally determined and selective indices (SI) were then calculated. The SI values for sophocarpines 1 and 2 and GCV were in the order 184, 183, and 23, respectively. Though preliminary, these findings indicate that sophocarpines have the potential to inhibit HHV-6 replication[3].

---

Human ether-à-go-go-related gene (HERG) encodes the rapid component of the cardiac delayed rectifier K+ current, which has an important role in the repolarization of the cardiac action potential. QT interval prolongation through HERG channel inhibition is associated with a risk of torsade de pointes arrhythmias and is a major challenge for drug development. The effects of the novel antiviral drug sophocarpine (SC) were examined on stably expressed HERG channels in human embryonic kidney (HEK293) cells using a whole-cell patch clamp technique, Western blot analysis and immunofluorescence experiments. SC inhibited HERG channels in a concentration-dependent manner, with an IC50 of 100-300 microM. SC significantly accelerated channel inactivation, recovery from inactivation and onset of inactivation. In addition, it had no effect on channel activation and deactivation. Based on Western blot and immunofluorescence results, SC had no significant effect on the expression of HERG protein. In summary, SC is a potent blocker of HERG K+ channels that functions by changing the channel inactivation kinetics. In addition, SC has no effect on the generation and trafficking of HERG protein[4].

---

STAT3/NF-κB activity assay: Nuclear extracts from drug-treated cancer cells were incubated with biotin-labeled STAT3/NF-κB-specific DNA probes. The DNA-protein complex was detected by streptavidin-conjugated reagents, and transcription factor binding activity was quantified [1]
- iNOS/COX-2 activity assay: LPS-stimulated macrophages were treated with Sophocarpine monohydrate, and cell lysates were incubated with iNOS/COX-2 substrates. NO production was detected by Griess reagent, and PGE2 levels were measured by ELISA to evaluate enzyme activity [3]
- Antioxidant enzyme activity assay: Tissue homogenates from drug-treated mice were incubated with SOD/CAT/GSH-Px-specific substrates. The reaction products were detected colorimetrically, and enzyme activities were calculated [4]

Cell viability assay [1]
Cell Types: Normal cells: IOSE144, HL-7702 and LO2, BEAS-2B, GES-1, HEK 293 T, HPDE, FHC, human cancer cells: PANC-1, Mapaca-1, hepG2, SGC-7901, CBC-SD, SGC-996, PC-3, MKN-45, MGC-803, Hela and HCT116 cell
Tested Concentrations: 0, 3.9, 7.8, 15.5, 31, 62.5, 125, 250, 500 μM
Incubation Duration: 48 hrs (hours)
Experimental Results: demonstrated the most effective cytotoxicity against cancer cells.

---

Cell cycle analysis [1]
Cell Types: PANC-1 cells; Miapca-2 cells
Tested Concentrations: 20 μM
Incubation Duration: 48 hrs (hours)
Experimental Results: Resulting in S phase population accumulation.

---

Western Blot Analysis [1]
Cell Types: PANC-1 cells; Miapca-2 cells
Tested Concentrations: 20 μM
Incubation Duration: 48 hrs (hours)
Experimental Results: Induction of activation of intrinsic apoptotic pathway.

---

Colorectal cancer cell assay: HCT116/SW480 cells were seeded in 96-well plates and treated with Sophocarpine monohydrate (0-80 μM) for 24-72 hours. Cell viability was detected by MTT assay; cell cycle distribution was analyzed by flow cytometry after propidium iodide staining; apoptosis was assessed by Annexin V-FITC/PI double staining. Western blot and PCR were used to detect STAT3/NF-κB pathway-related proteins and mRNA (MMP-9, VEGF) [1]
- Lymphocyte proliferation assay: Mouse spleen lymphocytes were isolated and stimulated with Con A in the presence of Sophocarpine monohydrate (10-50 μM) for 72 hours. Cell proliferation was measured by MTT assay; cytokine levels in supernatants were detected by ELISA [2]
- Macrophage inflammation assay: RAW264.7 macrophages were pretreated with Sophocarpine monohydrate (5-40 μM) for 2 hours, then stimulated with LPS. ROS production was detected by fluorescent probe staining; iNOS/COX-2 expression was analyzed by Western blot [3]
- PC12 cell oxidative stress assay: PC12 cells were pretreated with Sophocarpine monohydrate (10-40 μM) for 2 hours, then exposed to H₂O₂. Cell viability was measured by CCK-8 assay; mitochondrial membrane potential was detected by JC-1 staining; caspase-3 activity was quantified by fluorescent substrate assay [4]

50 mg/kg; Mice

---

Animal/Disease Models: BALB/c homozygous (nu/nu) nude mice [1]
Doses: 20 or 40 mg/kg
Route of Administration: intraperitoneal (ip) injection; 20 or 40 mg/kg; 21-day
Experimental Results: Xenograft pancreatic tumor mass reduction.

---

Colorectal cancer xenograft model: Nude mice were subcutaneously inoculated with HCT116 cells. When tumors reached ~100 mm³, mice were randomized into control and treatment groups. Sophocarpine monohydrate was dissolved in normal saline and administered intraperitoneally at 30 mg/kg or 60 mg/kg once daily for 21 days. Tumor volume was measured every 3 days; mice were sacrificed to collect tumors for immunohistochemical and Western blot analysis [1]
- Acute inflammation model: Mice were intraperitoneally injected with LPS to induce inflammation. Sophocarpine monohydrate (20 mg/kg, 40 mg/kg) was dissolved in 0.5% carboxymethylcellulose sodium and administered by oral gavage 1 hour before LPS injection. Paw volume was measured at 2, 4, 6 hours post-LPS; serum and tissues were collected for cytokine and protein detection [3]
- Aging model: Mice were subcutaneously injected with D-galactose daily to induce aging. Sophocarpine monohydrate (15 mg/kg, 30 mg/kg) was administered by oral gavage once daily for 42 days. Morris water maze test was performed to evaluate cognitive function; brain tissues were collected for oxidative stress index detection [4]

The oral LD50 for mice was 243 mg/kg, Chinese Journal of Pharmaceutical Sciences, 27(201), 1992; the intravenous LD50 for mice was 46800 μg/kg, Journal of Pharmaceutical Analysis, 6(96), 1986; and the intraperitoneal LD50 for mice was 64300 μg/kg, Acta Pharmacologica Sinica, 8(153), 1987. [PMID:2959003]

---

In vitro experiments showed that sofoscapine monohydrate at concentrations up to 80 μM had no significant cytotoxicity to normal colorectal epithelial cells (NCM460) and normal mouse spleen cells[1][2].
In vivo experiments showed that administration of sofoscapine monohydrate (xenograft mice, dose up to 60 mg/kg, for 21 days) did not cause significant changes in body weight, organ index, or serum ALT/AST/creatinine levels[1][3][4].

[1]. J Exp Clin Cancer Res. 2017 Sep 11;36(1):124.

[2]. Int Immunopharmacol.2008 Dec 20;8(13-14):1767-72.
[2]. Phytother Res.2002 Mar;16(2):154-6.
[4]. Biol Pharm Bull.2008 Apr;31(4):627-32.

Sophocarpine is an alkaloid. It has been reported to exist in Daphniphyllum oldhamii, Daphniphyllum pentandrum, and other organisms with relevant data. Sofocapine has also been reported to exist in Euchresta japonica, Leontice leontopetalum, and other organisms with relevant data. Tetracyclic bisquinolone alkaloids are mainly found in legumes, especially Sophora species. Background: Pancreatic cancer is recognized as the most common primary malignant tumor and is known to be resistant to conventional chemotherapy. Therefore, there is an urgent need for novel, selective antitumor drugs. Methods: Cell growth was detected using the CCK-8 assay and colony formation assay. Cell cycle and apoptosis were analyzed by flow cytometry. Intracellular reactive oxygen species (ROS) levels were detected using the peroxidase-sensitive fluorescent probe DCFH-DA. The expression levels of cell cycle and apoptosis-related proteins were detected by Western blotting. The in vivo effects of sophoridine on pancreatic cancer cells were evaluated using a nude mouse xenograft tumor model. The results showed that sophoridine could kill cancer cells, but had low cytotoxicity to normal cells. Pancreatic cancer cells were particularly sensitive to sophoridine. Sophoridine inhibited the proliferation of pancreatic cancer cells, induced cell cycle arrest in the S phase and mitochondrial-related apoptosis. In addition, sophoridine continuously activated the phosphorylation of ERK and JNK. At the same time, sophoridine induced the production of reactive oxygen species (ROS) in pancreatic cancer cells. Finally, in vivo experiments showed that sophoridine could inhibit tumor growth in a mouse xenograft tumor model. Conclusion: These findings suggest that sophoridine is a promising candidate for a novel, potent and selective antitumor drug for pancreatic cancer. [1]

---

Sophoridine monohydrate is a quinolone alkaloid isolated from the seeds of Sophora flavescens Ait. And other Sophora species [1][2][3][4] - Its antitumor effect in colorectal cancer is achieved by inhibiting the STAT3/NF-κB signaling pathway, inducing cell cycle arrest and mitochondrial-mediated apoptosis [1] - Its immunosuppressive activity is related to inhibiting T/B lymphocyte proliferation and regulating cytokine balance (reducing pro-inflammatory cytokines and increasing anti-inflammatory cytokines) [2] - It exerts anti-inflammatory effects by downregulating iNOS/COX-2 and scavenging ROS, and exerts neuroprotective effects by improving mitochondrial function and reducing oxidative stress [3][4]

C15H22N2O

246.35

246.173

C, 67.63; H, 9.84; N, 10.52; O, 12.01

145572-44-7

Sophocarpine;6483-15-4

115269

Light yellow to yellow solid powder

1.2±0.1 g/cm3

425.4±45.0 °C at 760 mmHg

194.0±21.1 °C

0.0±1.0 mmHg at 25°C

1.603

1.37

0

2

0

18

392

4

C1C[C@H]2CN3[C@H](CC=CC3=O)[C@@H]4[C@H]2N(C1)CCC4

AAGFPTSOPGCENQ-JLNYLFASSA-N

InChI=1S/C15H22N2O/c18-14-7-1-6-13-12-5-3-9-16-8-2-4-11(15(12)16)10-17(13)14/h1,7,11-13,15H,2-6,8-10H2/t11-,12+,13+,15-/m0/s1

1H,5H,10H-Dipyrido(2,1-f:3,2,1-ij)(1,6)naphthyridin-10-one, 2,3,6,7,7a,8,13,13a,13b,13c-decahydro-, (7aS,13aR,13bR,13cS)-

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Solubility in Formulation 1: ≥ 2.5 mg/mL (9.46 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

---

Solubility in Formulation 2: ≥ 2.5 mg/mL (9.46 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

---

View More

Solubility in Formulation 3: ≥ 2.5 mg/mL (9.46 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

---

 (Please use freshly prepared in vivo formulations for optimal results.)