(S)-10-Hydroxycamptothecin 19685-09-7_Topoisomerase_DNA Damage_Signaling Pathways_Products

10-Hydroxycamptothecin (also abbreviated as 10HCPT; 10OHCPT; HCPT; 10 HCPT), the 10-hydroxylated form of camptothecin, is a natural product and topoisomerase I inhibitor with potential anticancer activity. It significantly and dose-dependently inhibits HMEC migration, with an IC50 of 0.63 μM. The SRB assay was used to detect the proliferation of seven human tumor cell lines and human microvascular endothelial cells (HMEC). This led to an investigation into the antiangiogenic potential of 10-hydroxycamptothecin (HCPT), and two in vitro model systems were used to evaluate endothelial cell migration and tube formation.

Physicochemical Properties

Molecular Formula

C20H16N2O5

Molecular Weight

364.35

Exact Mass

364.105

Elemental Analysis

C, 65.93; H, 4.43; N, 7.69; O, 21.96

CAS #

19685-09-7

Related CAS #

19685-09-7

PubChem CID

97226

Appearance

Light yellow solid powder

Density

1.6±0.1 g/cm3

Boiling Point

820.7±65.0 °C at 760 mmHg

Melting Point

265-270°C

Flash Point

450.1±34.3 °C

Vapour Pressure

0.0±3.1 mmHg at 25°C

Index of Refraction

1.777

LogP

1.32

Hydrogen Bond Donor Count

Hydrogen Bond Acceptor Count

Rotatable Bond Count

Heavy Atom Count

Complexity

774

Defined Atom Stereocenter Count

SMILES

O1CC2C(N3CC4=CC5C=C(C=CC=5N=C4C3=CC=2[C@@](CC)(C1=O)O)O)=O

InChi Key

HAWSQZCWOQZXHI-FQEVSTJZSA-N

InChi Code

InChI=1S/C20H16N2O5/c1-2-20(26)14-7-16-17-11(5-10-6-12(23)3-4-15(10)21-17)8-22(16)18(24)13(14)9-27-19(20)25/h3-7,23,26H,2,8-9H2,1H3/t20-/m0/s1

Chemical Name

(19S)-19-ethyl-7,19-dihydroxy-17-oxa-3,13-diazapentacyclo[11.8.0.02,11.04,9.015,20]henicosa-1(21),2(11),3,5,7,9,15(20)-heptaene-14,18-dione

Synonyms

10-HCPT; Hydroxycamptothecin; 10OHCPT; HCPT; 10hydroxycamptothecine; 10HCPT; 10 HCPT; (S)10Hydroxycamptothecin; 10Hydroxy camptothecin; Hydroxycamptothecine; Camptothecin hydroxy;10hydroxycamptothecin

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

ln Vitro

10-Hydroxycamptothecin has a more potent inhibitory effect on BT-20 and MDA-231 cell growth than camptothecin (CPT), with IC50 values of >500 nM and 34.3 nM and 7.27 nM, respectively. 10-Hydroxycamptothecin exhibits >50-fold potency compared to CPT, which has an EC50 of 18.85 μM, by potently inducing human topoisomerase I-mediated cleavable complex formation of pBR322 plasmid DNA at 0.35 μM.[2] 10- Human microvascular endothelial cells (HMEC) treated with hydroxycamptothecin exhibit dose-dependent growth inhibition with an IC50 of 0.31 μM and significantly reduced HMEC migration with an IC50 of 0.63 μM. Additionally, treatment with 10-Hydroxycamptothecin inhibits tube formation in HMEC cells in a dose-dependent manner, with an IC50 of 0.96 μM.[3] 10-Hydroxycamptothecin (5–20 nM) dramatically suppresses Colo 205 cell growth, stops the cells in the G2 phase of the cell cycle, and triggers apoptosis via a mechanism that depends on caspase-3.[4]

ln Vivo

10-Hydroxycamptothecin treatment significantly reduces angiogenesis in the CAM model in a concentration-dependent manner, with 95% inhibition at a dose of 25 nM.This is more potent than suramin, which only inhibits angiogenesis by 60% at 125 nM.[3] Mice receiving oral dosages of 2.5–7.5 mg/kg of 10-hydroxycamptothecin every two days exhibit a notable reduction in the growth of Colo 205 xenografts without experiencing any acute toxicity. [4] LD50: 104 mg/kg (i.p.) in mice.[5]

Enzyme Assay

7-Alkyl, 7-alkyl-10-hydroxy, 7-alkyl-10-methoxy, and 7-alkyl-10, 11-methylenedioxy analogs of camptothecin have been synthesized and evaluated for their ability to trap human DNA topoisomerase I in cleavable complexes. The 7-alkyl chain lengths varied linearly from methyl to butyl. The concentration required to produce cleavable complexes with purified topoisomerase I in 50% of the plasmid DNA (EC(50)) was reduced by 1 order of magnitude by the introduction of a 10-methoxy or 7-alkyl group compared with camptothecin. The EC(50) values were reduced by 2 orders of magnitude with a 10-hydroxy or 10, 11-methylenedioxy moiety compared with camptothecin. The steady-state EC(50) concentrations for all of the analogs tested were slightly dependent on substitution at the 7-position, but this dependence was least with the 10-methoxy series. The kinetics of the reversibility of the complexes formed with all analogs was only slightly influenced by the length of the 7-substitution, with the trend that ethyl or greater lengths led to slightly reduced rate constants for cleavable complex reversal. These results were also observed for DNA-protein cross-link formation by the analogs in isolated CEM cell nuclei. Our data indicate that in vitro cleavable complex stability, as determined by the apparent rate constants for complex dissociation, does not reflect the in vitro biological activity of these camptothecin analogs. However, complex stability in vivo may be important for the antitumor activity of the compounds[3].

Cell Assay

In this study, researchers investigated the antiproliferative effect of 10-hydroxycamptothecin (HCPT), an alkaloid isolated from Camptotheca acuminata, on the human promyelocytic leukemia cell line, HL-60, and a 4'-(9-acridinylamino)methanesulfon-m-anisidide (m-AMSA)-resistant mutant, HL-60/m-AMSA. Using trypan blue dye exclusion and colony formation, doses of HCPT ranging from 0.01 to 1 microM progressively inhibited growth in both cell lines in a concentration-dependent manner. A minimal cross-resistance, approximately five-fold, between the wild-type and resistant cells was observed. Using the technique of alkaline elution, HCPT produced DNA single-strand breaks and protein-associated DNA strand cleavage in HL-60 and HL-60/m-AMSA cells. Quantitative analysis of drug-induced protein-DNA complexes was performed using sodium dodecyl sulfate-potassium chloride precipitation. In both cell lines, a good correlation with HCPT-induced cytotoxicity was observed. Similar results were achieved in wild-type cells treated with m-AMSA. Enzyme activity was measured in nuclei isolated from HL-60 and HL-60/m-AMSA cells, and in each case HCPT inhibited topoisomerase I activity to the same extent. The data suggest that the principle mechanisms for HCPT-induced cytotoxicity in HL-60 and HL-60/m-AMSA cells are inhibition of DNA topoisomerase I and production of protein-associated DNA strand breaks.[1]

For 72 hours, different concentrations of 10-Hydroxycamptothecin are applied to cells. The conventional MTT assay is used to track cell growth.

Animal Protocol

BALB/c-nu mice transplanted subcutaneously with Colo 205 cells
~7.5 mg/kg
Orally once per two days
BALB/c-nu mice transplanted subcutaneously with Colo 205 cells

Moreover, following oral administration at doses of 2.5-7.5 mg/kg/2 days, significant suppression of tumor growth by 10-HCPT was observed in mouse xenografts. No acute toxicity was observed after an oral challenge of 10-HCPT in BALB/c-nude mice every 2 days. The results of this study suggest that a relatively low dose of 10-HCPT (p.o.) is able to inhibit the growth of colon cancer, facilitating the development of a new protocol of human trials with this anticancer drug.[2]

Toxicity/Toxicokinetics

rat LD intravenous >10 mg/kg

References

[1]. Cancer Biochem Biophys . 1990 Jan;11(1):23-30.

[2]. Oncol Rep . 2006 May;15(5):1273-9.

[3]. Mol Pharmacol . 2000 Feb;57(2):243-51.

[4]. Life Sci . 2001 Aug 24;69(14):1619-28.

Additional Infomation

10-Hydroxycamptothecin is a pyranoindolizinoquinoline.
10-hydroxycamptothecin is under investigation in clinical trial NCT00956787 (Study of AR-67 (DB-67) in Myelodysplastic Syndrome (MDS)).
10-Hydroxycamptothecin has been reported in Nothapodytes nimmoniana, Fusarium solani, and Camptotheca acuminata with data available.

Solubility Data

Solubility (In Vitro)

DMSO: 8~50 mg/mL (22.0~137.2 mM)

Water: <1 mg/mL

Ethanol: <1 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: 2.5 mg/mL (6.86 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
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 (6.86 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication.
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.

Solubility in Formulation 3: 30% PEG400+0.5% Tween80+5% Propylene glycol : 30mg/mL

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

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.7446 mL	13.7231 mL	27.4461 mL
	5 mM	0.5489 mL	2.7446 mL	5.4892 mL
	10 mM	0.2745 mL	1.3723 mL	2.7446 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

(S)-10-Hydroxycamptothecin 19685-09-7

CAS No.: 19685-09-7

Physicochemical Properties

Biological Activity

Solubility Data