Alanosine

Chemical Properties

Crystalline Powder

Originator

Alanosine ,Triangle Pharmaceuticals

Uses

L-Alanosine is an antitumor antibiotic substance from the fermentation of Streptomyces alanosinicus. It is the first natural product found to have a N-nitrosohydroxylamino group on an aliphatic chain. It is also an experimental insect reproduction inhibitor.

Uses

Antibiotic substance from the fermentation of Streptomyces alanosinicus. The first natural product found to have a N-nitrosohydroxylamino group on an aliphatic chain. An experimental insect reproduction inhibitor

Manufacturing Process

Alanosine is an antibiotic isolated from the fermentation broth of Streptomyces alanosinicus n. sp.
Oat meal agar slants seeded with Streptomyces alanosinicus n. sp. ATCC 15710 were incubated at 20°C for 7 to 10 days and then used to inoculate 100 ml of a peptone-agar-glucose-yeast extract medium contained in 500 ml Erlenmeyer flask. The composition of this fermination medium is:
Meat extract 5.0 g./liter
Peptone 5.0 g./liter
Yeast extract 5.0 g./liter
Enzymatic casein hydrolysate 3.0 g./liter
Cerelose 2.0 g./liter
NaCl 1.5 g./liter
The medium is adjusted to pH 7.2 prior to sterilization for 20 minutes at 121°C and 15 Ibs steam pressure. The germination flasks are incubated at 28°C for 48 hours on rotary shaker having a 2 inch throw and making 240 rpm. A 3% transfer is made from the germination flask to 500 ml Erlenmeyer fermentation flaks containing 100 ml of medium TVF/5 having the following composition:
Glucose 50.0 g/L
Dried whale meat (Pascor) 10.0 g/L
CaCO4 5.0 g/L
(NH4)2SO4 1.0 g/L
MgSO4·7H2O 1.0 g/L
CuSO4 sol. 0.5% 1 ml
FeSO4 sol. 0.1% 1 ml
ZnSO4 sol. 0.2% 1 ml
MnSO4 sol. 0.8% 1 ml
The medium is adjusted to pH 7.0 prior to sterilization for 20 minutes at 121°C. The fermentation flasks are incubated and agitated under similar conditions as the germination flasks. After 72 hours the mycelium was separated by centrifugation and the untreated broth assayed by the streak dilution method. 30 liters of broth are centrifuged and 6% of Darco G-60 charcoal is added to the clear solution, which is stirred for 30 minutes; then the charcoal is filtered off to give an almost colorless solution which is concentrated in vacuum at 45-50°C to 1.5 liters. The concentrated solution is poured under stirring into 5 liters of methanol, the formed precipitate is filtered, washed with much acetone and dried in vacuum. Yield 230 g of crude product assaying about 10%.

Therapeutic Function

Antineoplastic

Enzyme inhibitor

This antibiotic and aspartate/asparagine analogue (FW = 149.11 g/mol; CAS 5854-93-3), also named L-2-amino-3-(N-hydroxy-N-nitrosoamino) propionic acid, isolated from fermentation broths of Streptomyces alanosinicus, inhibits adenylosuccinate synthetase, disrupting de novo synthesis of AMP in both malignant and normal cells. That alanosine inhibits the adenylosuccinate synthetase reaction, the first step in the conversion of IMP to AMP, is evidenced by the accumulation of IMP, but not adenylosuccinate, in treated cells. L-Alanosine’s action is potentiated in methylthioadenosine phosphorylase (MTAP) deficiency. Clinical use is limited by its toxicity. At a concentration as low as 2.7 μM, L-alanosine completely inhibits the incorporation of hypoxanthine into adenosine triphosphate by cultured Novikoff rat hepatoma cells. A related agent, L-alanosyl-5-aminoimidazole-4-carboxylic acid ribonucleotide (or alanosyl-AICOR) has been synthesized enzymatically using 4-(N-succino)- 5-aminoimidazole-4-carboxamide ribonucleotide (or SAICAR) synthetase in conjunction with 5-aminoimidazole-4-carboxylic acid ribonucleotide and alanosine. Alanosyl-AICOR is not a substrate of adenylosuccinate lyase from rat skeletal muscle, but it was an apparent competitive inhibitor in both reactions catalyzed by the enzyme.