Synonym name: Alkaloid V; 11-Deoxojervine
Catalogue No.: BP0437
Cas No.: 4449-51-8
Formula: C27H41NO2
Mol Weight: 411.63
Botanical Source: Veratrum californicum and Veratrum album (Liliaceae)
Purity: >99%
Analysis Method: HPLC-DAD or/and HPLC-ELSD
Identification Method: Mass, NMR
Packing: Brown vial or HDPE plastic bottle
Can be supplied from milligrams to grams, up to Kgs.
It can be produced in GMP plant on request.
Cyclopamine (11-deoxojervine) is a naturally occurring chemical that belongs to the group of steroidal jerveratrum alkaloids. It is a teratogen isolated from the corn lily (Veratrum californicum) that causes usually fatal birth defects. It can prevent the fetal brain from dividing into two lobes (holoprosencephaly) and cause the development of a single eye (cyclopia). Cyclopamine does so by inhibiting the hedgehog signaling pathway (Hh). Cyclopamine is useful in studying the role of Hh in normal development, and as a potential treatment for certain cancers in which Hh is overexpressed.
Cyclopamine was named for one-eyed lambs which were born to sheep which grazed on wild corn lily at a farm in Idaho. In 1957 the US Department of Agriculture started an eleven-year investigation which led to the identification of cyclopamine as the cause of the birth defect.
References
Storage conditions：Short-term storage at 2~8℃, long-term storage at -20 ~ -80℃
Cyclopamine is a natural plant derived steroid alkaloid that has attracted much attention for its significant role in regulating the Hedgehog (Hh) signaling pathway. The Hedgehog signaling pathway, as a key regulatory mechanism for embryonic development and tissue homeostasis maintenance, is closely related to the occurrence and development of various tumors, especially in malignant tumors such as basal cell carcinoma (BCC). As a selective SMOothened (SMO) receptor inhibitor, cyclophosphamide can effectively block the abnormal activation of the Hh signaling pathway, demonstrating potential anti-tumor activity.
This article aims to provide a systematic review of the chemical structure and physicochemical properties, plant sources and extraction methods, pharmacological activity and mechanism of action, pharmacological evaluation, pharmacokinetic characteristics, as well as the prospects and challenges of its clinical application in the field of natural product pharmacology and tumor targeted therapy. The goal is to provide scientific reference and theoretical support for this field.
The molecular formula of cyclohexylamine is C27H41NO2, with a molecular weight of 411.62 and a CAS number of 4449-51-8. Its structure belongs to steroid alkaloids, with a typical steroid skeleton containing a nitrogen-containing cyclic structure that endows it with unique biological activity. The LogP value of cyclophosphamide is 4.5, indicating its high lipid solubility, which is beneficial for membrane penetration and in vivo distribution. The polar surface area (TPSA) is 38.69 Å ², and the number of hydrogen bond acceptors is 3, indicating that its molecular polarity is moderate and conducive to binding with target proteins.
The physicochemical properties of cyclophosphamide enable it to effectively cross the blood-brain barrier (BBB), which has potential advantages for the treatment of central nervous system related diseases. In addition, cyclophosphamide showed low risks of hepatotoxicity and cardiotoxicity in in vitro experiments, and did not exhibit hERG channel inhibition. The Ames mutagenicity test results were negative, indicating its high safety and good pharmacological basis.
Cyclophosphamide was first isolated from the wild plant Veratrum californicum, native to North America. This plant belongs to the Liliaceae family and has traditionally been used as a folk herb to treat various diseases. The discovery of cyclophosphamide originated from the study of sheep fetal malformation (sheep head deformity) caused by this plant, revealing its interference with the Hh signaling pathway during embryonic development.
The extraction of cyclohexylamine is usually carried out using organic solvent extraction combined with column chromatography separation technology. The specific process includes:
1. Collect dried aboveground parts or rhizomes of Veratrum californicum and crush them.
2. Reflux extraction is carried out using methanol or ethanol, and the extract is concentrated and then subjected to solvent distribution to remove impurities.
3. Further purify by silica gel column chromatography or high performance liquid chromatography (HPLC) to obtain high-purity cyclohexylamine.
In recent years, with the development of molecular biology and chemical synthesis technology, the semi synthesis and total synthesis methods of cyclopamine have gradually matured, providing technical support for its large-scale production and structural optimization.
As a specific inhibitor of the Hedgehog signaling pathway, cyclophosphamide's pharmacological activity mainly manifests in its antagonistic effect on SMO receptors. Cell experiment data shows that the inhibitory IC50 of cyclophosphamide on SMO is approximately 46 nM, demonstrating efficient targeting activity. By blocking SMO, cyclophosphamide effectively inhibits the activation of downstream transcription factor GLI, thereby downregulating the expression of genes related to the Hh pathway.
In a variety of tumor models, cycloparamide has shown significant anti proliferation and pro apoptosis effects, especially in basal cell carcinoma, pancreatic cancer and some brain tumors. Animal experiments have shown that cyclophosphamide can significantly slow down tumor growth and reduce tumor burden. In addition, cyclophosphamide has low toxicity to normal cells and exhibits good selectivity.
In addition to its anti-tumor effect, the application of cyclophosphamide in embryonic developmental abnormalities models also reveals its unique value in regulating the Hh signaling pathway, providing an important tool for studying developmental biology and congenital disease mechanisms.
The Hedgehog signaling pathway plays a crucial role in embryonic development and tissue homeostasis, with its core components including ligand Hh protein, receptor PTCH1, signal transduction protein SMO, and transcription factor GLI family. Under normal circumstances, PTCH1 inhibits SMO activity and blocks signal transduction; After binding to PTCH1 with Hh ligand, the inhibition of SMO is released and downstream signals are activated.
Cyclophosphamide directly binds to SMO, blocking its activation and inhibiting the transmission of the Hh signaling pathway. Its main targets include:
- SMO（Smoothened）Cyclobalamin acts as a selective inhibitor of SMO, blocking its conformational changes and signal transduction.
- PTCH1（Patched1）As a negative regulatory receptor of the Hh pathway, dysfunction of PTCH1 often leads to abnormal activation of SMO, and cyclophosphamide indirectly regulates the signal feedback of PTCH1 by inhibiting SMO.
- GLI1（Glioma-associated oncogene homolog 1）As the main transcription factor of the Hh pathway, the expression and activity of GLI1 are regulated by SMO, and cyclophosphamide reduces the transcriptional activity of GLI1 by inhibiting SMO.
- SUFU (Suppressor of Fused) and HHIP (Hedgehog interacting protein)The effect of cyclohexylamine on these regulatory proteins is still in the research stage, but its overall regulatory network is crucial for maintaining the homeostasis of the Hh signaling pathway.
The molecular binding mechanism of cyclohexylamine has been partially elucidated through crystallographic and computational simulations, showing its binding to the seven transmembrane domains of SMO receptors, preventing the activation conformation of SMO and thereby inhibiting signal transmission.
The pharmacological parameters of cyclophosphamide indicate that it has good potential for drug development. Its molecular weight (411.62) and LogP (4.5) meet the basic requirements for oral active drugs in Lipinski's rules, and moderate polarity and hydrogen bond receptor number are beneficial for target binding and bioavailability.
Cyclophosphamide can effectively cross the blood-brain barrier, indicating its potential application in central nervous system diseases. The in vivo toxicological evaluation showed that cyclophosphamide had no significant hepatotoxicity or cardiotoxicity, and the hERG channel inhibition experiment was negative, reducing the risk of arrhythmia. The Ames mutagenicity test result is negative, indicating a low risk of genetic toxicity.
Pharmacokinetic studies have shown that cyclophosphamide is well absorbed orally, has a moderate plasma half-life, and is widely distributed in the body. Its metabolism is mainly carried out through the liver cytochrome P450 enzyme system, with stable metabolites and excretion mainly through bile and urine. The drug interaction risk of cyclophosphamide is low, but attention should be paid to potential competition with other CYP450 substrates.
As one of the first discovered natural Hedgehog signaling pathway inhibitors, cyclophosphamide provides a new strategy for targeted therapy of basal cell carcinoma and related tumors. The abnormal activation of SMO in basal cell carcinoma is the core of its pathogenesis, and cyclophosphamide significantly inhibits tumor cell proliferation and invasion by selectively inhibiting SMO. The preclinical research results are encouraging.
At present, cycloparamide and its derivatives have entered a number of clinical trials to evaluate their efficacy and safety in basal cell carcinoma, pancreatic cancer, glioma and other tumors. Some analogues of cyclohexylamine, such as Vismodegib, have been approved by the FDA for the treatment of advanced basal cell carcinoma, demonstrating the clinical value of Hh pathway inhibitors.
Future research directions include:
-Optimize the pharmacokinetic properties of cyclophosphamide, improve bioavailability and tissue selectivity.
-Reducing drug resistance through structural modification and overcoming drug resistance in tumor cells.
-Explore the application of cyclophosphamide in other diseases related to Hh signal abnormalities, such as fibrosis, autoimmune diseases, etc.
-Combining other targeted drugs or immunotherapy to enhance anti-tumor efficacy and improve patient prognosis.
As a natural Hedgehog signaling pathway inhibitor, cyclopamine has shown broad application prospects in the field of tumor targeted therapy due to its unique chemical structure and significant biological activity. Its selective antagonistic effect on SMO receptors provides new ideas and drug basis for the treatment of basal cell carcinoma and related malignant tumors. Despite facing challenges in pharmacokinetic optimization and drug resistance, the research progress of cyclophosphamide and its derivatives continues to drive the development of natural product pharmacology and precision medicine.
In the future, by combining modern drug design and molecular biology techniques, cyclophosphamide is expected to become an important drug for the treatment of Hedgehog signaling pathway related diseases, bringing more benefits to patients.
