Camptothecin (Campathecin) 7689-03-4_Topoisomerase_DNA Damage_Signaling Pathways_Products

Camptothecin (also known as Campathecin; CPT; NSC-100880), a naturally occurring quinoline alkaloid, is a potent and specific inhibitor of DNA enzyme topoisomerase I (Topo I) with potent antitumor activity. In a cell-free assay, it inhibits topoisomerase I (Topo I) with an IC50 of 0.68 μM. The bark and stem of Camptotheca acuminata can be used to isolate camptothecin, which is licensed for use in Traditional Chinese Medicine as an anticancer medication. In initial clinical trials, it demonstrated impressive anticancer activity; however, it also had low solubility and adverse drug reaction. Due to these drawbacks, medicinal and synthetic chemists have successfully created a number of syntheses of camptothecin and its derivatives in an effort to increase the chemical's advantages.

Physicochemical Properties

Molecular Formula

C20H16N2O4

Molecular Weight

348.35

Exact Mass

348.11

Elemental Analysis

C, 68.96; H, 4.63; N, 8.04; O, 18.37

CAS #

7689-03-4

Related CAS #

7689-03-4; 25387-67-1 (sodium)

PubChem CID

24360

Appearance

Light yellow solid powder

Density

1.5±0.1 g/cm3

Boiling Point

757.0±60.0 °C at 760 mmHg

Melting Point

260 °C (dec.)(lit.)

Flash Point

411.6±32.9 °C

Vapour Pressure

0.0±2.7 mmHg at 25°C

Index of Refraction

1.746

LogP

1.6

Hydrogen Bond Donor Count

Hydrogen Bond Acceptor Count

Rotatable Bond Count

Heavy Atom Count

Complexity

742

Defined Atom Stereocenter Count

SMILES

O1C([C@](C([H])([H])C([H])([H])[H])(C2C([H])=C3C4C(=C([H])C5=C([H])C([H])=C([H])C([H])=C5N=4)C([H])([H])N3C(C=2C1([H])[H])=O)O[H])=O

InChi Key

VSJKWCGYPAHWDS-FQEVSTJZSA-N

InChi Code

InChI=1S/C20H16N2O4/c1-2-20(25)14-8-16-17-12(7-11-5-3-4-6-15(11)21-17)9-22(16)18(23)13(14)10-26-19(20)24/h3-8,25H,2,9-10H2,1H3/t20-/m0/s1

Chemical Name

(19S)-19-ethyl-19-hydroxy-17-oxa-3,13-diazapentacyclo[11.8.0.02,11.04,9.015,20]henicosa-1(21),2,4,6,8,10,15(20)-heptaene-14,18-dione

Synonyms

Camptothecin; NSC-100880; (S)-(+)-Camptothecin; (S)-(+)-Camptothecin; NSC100880; Camptothecine; NSC 100880; d-Camptothecin; 20(S)-Camptothecine; (+)-Camptothecin; CPT; Camptothecine; (S)-(+)-Camptothecin; Campathecin; (+)-Camptothecin; (+)-Camptothecine; d-Camptothecin;

HS Tariff Code

2934.99.9001

Storage

Powder-20°C 3 years

4°C 2 years

In solvent -80°C 6 months

-20°C 1 month

Note: This product requires protection from light (avoid light exposure) during transportation and storage.

Shipping Condition

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

Biological Activity

Targets

Topoisomerase I ( IC50 = 679 nM ); Camptothecins

ln Vitro

Camptothecin, a plant alkaloid, was first isolated from Camptotheca acuminate in 1966.[1] It has been observed that camptothecin stops cells in their tracks during the S phase of mitosis. With IC50 values ranging from 37 nM to 48 nM, camptothecin demonstrates nanomolar potency in cytotoxicity against numerous human tumor cell lines, including HT29, LOX, SKOV3, and SKVLB.[2] At an IC50 of 13 μM, camptothecin causes apoptosis in primary mouse hepatocytes when combined with TNF. Moreover, camptothecin inhibited the expression of TNF-receptor associated factor 2 (TRAF2), X-linked inhibitor of apoptosis protein (X-IAP), and FLICE-inhibitory protein (FLIP), as well as the TNF-induced NF-κB Activation.[4] In HCT116 cells, camptothecin (5 μM) causes the mixed lineage leukemia 5 (MLL5) protein to be broken down by proteasomes, which phosphorylates p53 at Ser392. [5] Owing to camptothecin's poor solubility and unfavorable effects, a number of camptothecin analogues have been created; two of these, topotecan and irinotecan, have FDA approval and are used in cancer chemotherapy.

ln Vivo

Camptothecin (8 mg/kg) exhibits total growth inhibition and regression in mice given xenografts of different tumors, such as tumors of the ovaries, stomach, colon, lung, and breast.[3] Liver damage is induced in mice by combinations of 50 mg/kg Camptothecin and TNF (5 and 7 μg/kg), but not by Camptothecin alone.[4]

Enzyme Assay

The calf thymus is the source of topoisomerase I, which is absent from topoisomerase II. Every reaction is conducted in microtiter plates using 10 mL volumes of reaction buffer (50 mM Tris-HCl, pH 7.5, 100 mM KCl, 0.5 mM EDTA, and 30 pg/mL BSA). In 96-well microtiter plates, topoisomerase enzyme and 32P end-labeled pBR322 DNA are added after 10 mg/mL of camptothecin has been dissolved in DMSO and serially diluted. After the reaction mixture is incubated for 30 minutes at room temperature, 2 mL of a solution containing proteinase K and sodium dodecyl sulfate (1.6% and 0.14 mg/mL final concentrations, respectively) is added to stop the reaction. The samples are electrophoresed in 1.5% agarose gels in TBE buffer after the plates are heated to 50 °C for 30 minutes and 10 mL of standard stop mixture containing 0.45 N NaOH is added. Gels are exposed to X-ray film after being blotted on nitrocellulose paper, dried, and polished. The log drug concentration is plotted against the units of cleavage, which are computed from the autoradiographs. We then obtain the IC50 values.

Cell Assay

In 96-well microtiter plates, tumor cells are plated at a density of 1500–4000 cells per well in 100 μL of medium, and they are left to adhere for the entire night. After 48 hours of camptothecin incubation, cells are incubated for an additional 48 hours in fresh medium. Quadruplicates of each concentration of camptothecin are added. After treating the cells with MTT for four hours, the reduced dye product is removed from the cells using 0.2 mL of DMSO and then 50 μL of Sorensen's buffer. After giving the plates a quick shake, the absorbance at 570 nm is measured and recorded. An MTT assay data set is fitted with curves using a four-parameter logistic equation.

Animal Protocol

In vivo, CPT treatment sensitized mice to TNF-induced liver damage. In conclusion, the combination of topoisomerase inhibition and TNF blocks survival signaling and elicits a type of hepatocyte death similar to actinomycin D/TNF or galactosamine/TNF. During antitumor treatment with topoisomerase inhibitors, an impaired immune function often results in opportunistic infections, a situation where the systemic presence of TNF might be critical for the hepatotoxicity reported in clinical topoisomerase inhibitor studies.[4]

Nude mice (NIH-I high fertility strain) bearing xenografts of CASE, SW48, DOY, SPA, and CLO cells; 0–8 mg/kg; Administered via i.m. or i.v. injection

Nude mice (NIH-I high fertility strain) bearing xenografts of CASE, SW48, DOY, SPA, and CLO cells

References

[1]. J Am Chem Soc, 1966, 88 (16), 3888–3890.

[2]. J Med Chem . 1995 Feb 3;38(3):395-401.

[3]. Cancer Res . 1991 Jun 1;51(11):3052-5.

[4]. Hepatology . 2004 May;39(5):1311-20.

[5]. Oncogene, 2011, 30(33), 3599-3611.

Additional Infomation

Camptothecin is a pyranoindolizinoquinoline that is pyrano[3',4':6,7]indolizino[1,2-b]quinoline which is substituted by oxo groups at positions 3 and 14, and by an ethyl group and a hydroxy group at position 4 (the S enantiomer). It has a role as an EC 5.99.1.2 (DNA topoisomerase) inhibitor, an antineoplastic agent, a genotoxin and a plant metabolite. It is a pyranoindolizinoquinoline, a tertiary alcohol, a delta-lactone and a quinoline alkaloid.
Camptothecin is an alkaloid isolated from the stem wood of the Chinese tree, Camptotheca acuminata. This compound selectively inhibits the nuclear enzyme DNA topoisomerase, type I. Several semisynthetic analogs of camptothecin have demonstrated antitumor activity.
Camptothecin has been reported in Ophiorrhiza liukiuensis, Nothapodytes nimmoniana, and other organisms with data available.
Camptothecin is an alkaloid isolated from the Chinese tree Camptotheca acuminata, with antineoplastic activity. During the S phase of the cell cycle, camptothecin selectively stabilizes topoisomerase I-DNA covalent complexes, thereby inhibiting religation of topoisomerase I-mediated single-strand DNA breaks and producing potentially lethal double-strand DNA breaks when encountered by the DNA replication machinery. (NCI)
An alkaloid isolated from the stem wood of the Chinese tree, Camptotheca acuminata. This compound selectively inhibits the nuclear enzyme DNA TOPOISOMERASES, TYPE I. Several semisynthetic analogs of camptothecin have demonstrated antitumor activity.

Drug Indication
Investigated for the treatment of cancer.

Mechanism of Action
Camptothecin binds to the topoisomerase I and DNA complex resulting in a ternary complex, stabilizing it and preventing DNA re-ligation and therefore causes DNA damage which results in apoptosis.

Solubility Data

Solubility (In Vitro)

DMSO: 3~6.3 mg/mL (8.6~17.9 mM)

Water: <1 mg/mL

Ethanol: <1 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: 10 mg/mL (28.71 mM) in 15% Cremophor EL + 85% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

Solubility in Formulation 2: 30% PEG400+0.5% Tween80+5% Propylene glycol : 30 mg/mL

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

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.8707 mL	14.3534 mL	28.7068 mL
	5 mM	0.5741 mL	2.8707 mL	5.7414 mL
	10 mM	0.2871 mL	1.4353 mL	2.8707 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.

PeptideDB

Camptothecin (Campathecin) 7689-03-4

CAS No.: 7689-03-4

Physicochemical Properties

Biological Activity

Solubility Data