Fumagillin

Description

Fumagillin is a fungal metabolite that has been found in A. fumigatus and has diverse biological activities. It inhibits methionyl aminopeptidase 2 (METAP2; IC₅₀ = 10 nM). Fumagillin (10 ng/ml) inhibits tube formation in a rat blood vessel organ culture assay. It inhibits E. cuniculi replication in isolated rabbit kidney cells and canine embryo cells when used at a concentration of 5 μg/ml. In vivo, fumagillin (30 mg/kg per day) decreases tumor growth and the number of metastases in a mouse model of diethylnitrosamine-induced hepatocellular carcinoma. It also reduces subcutaneous and gonadal fat mass in a mouse model of high-fat diet-induced obesity. Formulations containing fumagillin have been used to treat conjunctival and intestinal microsporidial infections in immunocompromised patients.

Chemical properties

White powder

Originator

Fugillin,Upjohn,US,1953

The Uses of Fumagillin

Fumagillin is a compound isolated from the fungus Aspergillus fumigatus. Fumagillin is an antimicrobial agent used in the treatment of microsporidiosis. Fumagillin shows promise as both an an anti-inf ective and antiangiogenic agent.

The Uses of Fumagillin

Methionine aminopeptidase 2 inhibitor

The Uses of Fumagillin

Fumagillin is a polyene mycotoxin isolated from Aspergillus fumigatus in 1951 as a potent antiprotozoan for the treatment of amoebiasis. More recently, fumagillin has been shown to inhibit endothelial cell proliferation and angiogenesis by inhibiting methionine aminopeptidase-2 (MetAP-2).

What are the applications of Application

Fumagillin is an angiogenesis suppressing antibiotic and MetAP-2 inhibitor

Definition

An antibiotic substance produced by Aspergillus fumigatus.

Manufacturing Process

A fermentation medium comprising 4,600 gal of sterile corn steep-glucosecalcium carbonate medium in a 6,000-gal fermentation tank is adjusted to pH 6.0 with sodium carbonate prior to sterilization and thereafter inoculated with 200 gal of vegetative inoculum of Aspergillus fumigatus NRRL 2436. The inoculated medium is incubated for approximately 108 hours at a temperature of 26°C and agitated by an impeller rotating at 114 rpm and aerated at a rate of 500 cfm. An antifoam agent of the type used in penicillin fermentation is used as required.
The clarified liquid obtained from the fermentation medium (beer) by filtration in any of the usual apparatus for removing mycelia and suspended solids from fermentation beers, after first adjusting the pH of the contents of the fermentation tank to above about pH 7.0 and preferably to between pH 7.5 and pH 8.5 with, for example, the addition of an alkaline material such as sodium carbonate, is intimately mixed with hexane with a Podbielniak extractor and the hexane layer containing undesirable fatty material discarded. The pH of the defatted liquid is adjusted to about pH 3 by the addition of H2SO4, and the defatted liquid is extracted with chloroform. The chloroform is removed under reduced pressure without external heating. After the removal of all of the chloroform the residual syrup is dissolved in acetone. The acetone solution is cooled to 5°C whereupon a small quantity of brown precipitate separates which is removed by filtration. The precipitate is washed with acetone and the washings added to the original filtrate. A portion of the above acetone solution is concentrated under reduced pressure at room temperature under an atmosphere of nitrogen. The resulting thick suspension is placed in a 1-liter centrifuge cup, under nitrogen, and cooled at 30°C for 18 hours. The suspension is centrifuged for 1 hour at 1,500 to 1,700 rpm. The supernatant liquid is decanted from the residual solids which are washed 5 times at room temperature with several portions of tert-butanol. A residual solid material remains after the wash and after drying at room temperature. This material, after recrystallization from a mixture of equal parts of water and of methanol has a MP of 190°C to 192°C.

Therapeutic Function

Antibiotic

General Description

Fumagillin is an antibiotic obtained by the fermentation of certain strains of Aspergillus fumigatus with potent amoebicidal action.

Biological Activity

Antibiotic and antiangiogenic agent; covalently binds and inhibits methionine aminopeptidase-2. Inhibits endothelial cell proliferation in vitro and tumor-induced angiogenesis in vivo . Also inhibits tumor growth in mice. Analog available, TNP 470 (N-(2-Chloroacetyl)carbamic acid (3R,4S,5S,6R)-5-Methoxy-4-[(2R,3R)-2-methyl-3-(3-methyl -2-buten-1-yl)-2-oxiranyl]-1-oxaspiro[2.5]oct-6-yl ester).

storage

+4°C

Purification Methods

Forty grams of a commercial sample containing 42% fumagillin, 45% sucrose, 10% antifoam agent and 3% of other impurities are digested with 150mL of CHCl3. The insoluble sucrose is filtered off and washed with CHCl3. The combined CHCl3 extracts are evaporated almost to dryness at room temperature under reduced pressure. The residue is triturated with 20mL of MeOH, and the fumagillin is filtered off by suction. It is crystallised twice from 500mL of hot MeOH by standing overnight in a refrigerator (yellow needles). (The long-chain fatty ester used as antifoam agent is still present, but is then removed by repeated digestion, on a steam bath, with 100mL of diethyl ether.) For further purification, the fumagillin (10g) is dissolved in 150mL of 0.2M ammonia, and the insoluble residue is filtered off. The ammonia solution (cooled in running cold water) is then brought to pH 4 by careful addition of M HCl with constant shaking in the presence of 150mL of CHCl3. (Fumagillin is acid-labile and must be removed rapidly from the aqueous acid solution.) The CHCl3 extract is washed several times with distilled water, dried (Na2SO4) and evaporated under reduced pressure. The solid residue is washed with 20mL of MeOH. The fumagillin is filtered off by suction, then crystallised from 200mL of hot MeOH. [Tarbell et al. J Am Chem Soc 77 5610 1955.] Alternatively, 10g of fumagillin in 100mL CHCl3 is passed through a silica gel (5g) column to remove tarry material, and the CHCl3 is evaporated to leave an oil which gives fumagillin on crystallisation from amyl acetate. It recrystallises from MeOH (charcoal) or Me2CO/MeOH. The fumagillin is stored in dark bottles in the absence of oxygen and at low temperatures. [Schenk et al. J Am Chem Soc 77 5606 1955, Beilstein 19 III/IV 1012.]

References

1) Sin et al. (1997) The anti-angiogenic agent fumagillin covalently binds and inhibits the methionine aminopeptidase, MetAP-2; Proc. Natl. Acad. Sci. USA, 94 6099 2) Ingber et al. (1990) Synthetic analogues of fumagillin that inhibit angiogenesis and suppress tumour growth; Nature, 348 555