Abbott Laboratories Corporation

Ronald E Carney

James B McAlpine

Jerry R Martin

John S Tadanier

Robert L Devault

Arthur C Sinclair

Stephen Hanessian

Thomas John Perun

Ellen Marie Schwartz

Joseph Arndt

Julie R List

The present invention involves a process of purifying and recovering a recombinant protein from a suspension of cells. The process of the present invention involves extracting a recombinant protein from a concentrated suspension of cells using a water-miscible organic solvent, isolating the recombinant protein from the suspension of cells, concentrating the recombinant protein to remove the water-miscible organic solvent, precipitating the recombinant protein using an acid, washing the recombinant protein with the salt or free form of a suitable organic acid and recovering the recombinant protein.

Process for the recovery and purification of a recombinant protein from

Disclosed herein are fortimicin derivatives represented by the formula: ##STR1## wherein R.sub.z is a monocyclicaryloxycarbonyl amine protecting group, or is loweralkyl, hydroxyloweralkyl, loweracyl, hydroxyloweracyl, or a monocyclicaryloxycarbonyl-protected aminoloweralkyl, diaminoloweralkyl, N-loweralkylaminoloweralkyl, N,N-diloweralkylaminoloweralkyl, aminohydroxyloweralkyl, N-loweralkylaminohydroxy-loweralkyl, N,N-diloweralkylaminohydroxyloweralkyl, aminoloweracyl, diaminoloweracyl, N-loweralkylaminoloweracyl, N,N-diloweralkylaminoloweracyl, or aminohydroxyloweracyl; R.sub.1 is hydroxy or loweracyloxy; R.sub.2 is hydrogen, hydroxy or --OR.sub.4, wherein R.sub.4 is tert-butyldimethylsilyl or thiocarbonylimidazoyl; or R.sub.1 and R.sub.2 can be taken together to form ##STR2## wherein R.sub.5 and R.sub.6 are loweralkyl; R.sub.3 is hydroxy or loweracyloxy; and z is a monocyclicaryloxycarbonyl amine protecting group. The compounds are useful as intermediates in the preparation of 3-demethoxyfortimicins.

Intermediates for the preparation of 3-demethoxyfortimicins

An improved method of producing 2-epi-fortimicin A directly from fortimicin A by protecting the amine functions of fortimicin A, converting the N-protected fortimicin A to an intermediate of the formula: ##STR1## wherein L is a leaving group and Z is an amine-protecting group, reacting the intermediate (I) with a loweralkyl metal halide in N,N-dimethylformamide to form N-protected-2-epi-fortimicin A, and then hydrogenating the N-protected-2-epi-fortimicin A to obtain 2-epi-fortimicin A.

Method of producing 2-epi-fortimicin A

1-Epi-2-deoxyfortimicin B and 4-N-derivatives thereof represented by the formula: ##STR1## wherein: R.sub.1 is hydrogen or loweralkyl and R.sub.2 is selected from the group consisting of loweralkyl, aminoloweralkyl, diaminoloweralkyl, N-loweralkylaminoloweralkyl, N,N-diloweralkylaminoloweralkyl, hydroxyloweralkyl, N-loweralkylaminohydroxyloweralkyl, N,N-diloweralkylaminoloweralkyl, acyl, aminoacyl, diaminoacyl, hydroxyacyl, hydroxy-substituted aminoacyl hydroxy-substituted diaminoacyl, N-loweralkylaminoacyl, N,N-diloweralkylaminoacyl, hydroxy-substituted-N-loweralkylaminoacyl or hydroxysubstituted N,N-diloweralkylaminoacyl with the limitation that R.sub.2 cannot be hydrogen glycyl, formylglycyl or hydantoyl, and the pharmaceutically acceptable salts thereof. The compounds are broad spectrum antibiotics.

1-Epi-2-deoxyfortimicin B and derivatives

A method of producing 1-epi-2-deoxyfortimicin A, key intermediates therefor and an improved process for synthesizing the key intermediate 1,2-di-epi-fortimicin A.

Method of producing 1-epi-2-deoxyfortimicin A and intermediates therefor

Derivatives of O-demethylseldomycin factor 5 represented by the following formula are provided: ##STR1## wherein: R is hydrogen or loweralkyl; R.sub.1 and R.sub.2 can be either hydrogen or hydroxy with the limitation that both R.sub.1 and R.sub.2 cannot be hydroxy; R.sub.3 is selected from the group consisting of hydrogen, loweralkyl, aminoloweralkyl, diaminoloweralkyl, hydroxyloweralkyl, N-loweralkylaminoloweralkyl, aminohydroxyloweralkyl, N-loweralkylaminohydroxyloweralkyl, or ##STR2## with the limitation that when R.sub.1 is hydroxy, R.sub.3 cannot be hydrogen when R is hydrogen; and R.sub.4 is selected from the group consisting of loweralkyl, aminoloweralkyl, diaminoloweralkyl, hydroxyloweralkyl, N-loweralkylaminoloweralkyl, aminohydroxyloweralkyl and N-loweralkylaminohydroxyloweralkyl; and the pharmaceutically acceptable salts thereof. The compounds are potent anti-bacterial agents.

O-Demethylseldomycin factor 5 derivatives

A 1-Epi fortimicin represented by the formula ##STR1## wherein: R.sub.1 is hydrogen or loweralkyl; R.sub.2 is hydrogen or hydroxyl, R.sub.3 is selected from the group consisting of hydrogen, loweralkyl, aminoloweralkyl, diaminoloweralkyl, N-loweralkylaminoloweralkyl, N,N-diloweralkylaminoloweralkyl, hydroxyloweralkyl, aminohydroxyloweralkyl, N-loweralkylaminohydroxyloweralkyl, N,N-diloweralkylaminohydroxyloweralkyl, acyl, aminoacyl, diaminoacyl, hydroxyacyl, hydroxy-substituted aminoacyl, hydroxy-substituted diaminoacyl, N-loweralkylaminoacyl, N,N-diloweralkylaminoacyl, hydroxy-substituted N-loweralkylaminoacyl or hydroxy-substituted N,N-diloweralkylaminoacyl; R.sub.4 is hydrogen or methyl; and the pharmaceutically acceptable salts thereof.

1-Epi-fortimicin A and derivatives

A new seldomycin factor 5 derivative is provided, O-demethylseldomycin factor 5. The compound is represented by the formula: ##STR1## The compound is a potent anti-bacterial agent.

O-Demethylseldomycin factor 5

4,N, 2'-N and 4,2'-Di-N-derivatives of fortimicin AM and fortimicin AP are provided by this invention. The compounds are useful as anti-bacterial agents.

Fortimicins AM and AP derivatives

Disclosed are compounds of the formula ##STR1## wherein R is loweralkyl, aminoloweralkyl, hydroxyloweralkyl and aminohydroxyloweralkyl and the pharmaceutically acceptable acid addition salts thereof, all of which are potent antibacterials.

1-N-Substituted derivatives of seldomycin factor 5

Novel fortimicins, fortimicin AM and fortimicin AP are represented by the formula ##STR1## wherein R.sub.1 is hydrogen or hydroxy and R.sub.2 is hydrogen when R.sub.1 is hydroxy and hydroxy when R.sub.1 is hydrogen. Fortimicins AM and AP are coproduced with fortimicin A, fortimicin B and other minor factors in the fermentation of Micromonospora olivoasterospora. The compounds are useful as intermediates for 4-N-substituted fortimicins AP and AM which are useful as antibacterial agents.

Fortimicins AM and AP

5-Deoxyfortimicin A, 2,5-dideoxyfortimicin A and the corresponding 4-N-acyl and alkyl fortimicin B derivatives thereof, pharmaceutically acceptable salts thereof, intermediates therefor, and pharmaceutical compositions containing the compounds of this invention. The fortimicin derivatives are represented by the formula: ##STR1## wherein R is selected from the group consisting of acyl, hydroxyacyl, aminoacyl, N-monoloweralkylaminoacyl, N,N-diloweralkylaminoacyl, hydroxyacyl, hydroxy-substituted aminoacyl or an amino acid residue other than glycyl, loweralkyl, aminoloweralkyl, hydroxyloweralkyl, N-monoloweralkylaminoloweralkyl, N-N-diloweralkylaminoloweralkyl or hydroxy-substituted aminoloweralkyl; R.sub.1 is hydrogen or hydroxy; and R.sub.2 is hydrogen or hydroxy with the limitation that either R.sub.1 and R.sub.2 each are hydrogen or when only one of R.sub.1 or R.sub.2 are hydrogen, R.sub.2 is hydrogen and R.sub.1 is hydroxy and R.sub.1 and R.sub.2 both cannot be hydroxy.

5-Deoxyfortimicin A, 2,5-dideoxyfortimicin A and the corresponding

An improved method for producing the antibiotic 2-deoxyfortimicin A, the method comprising the process of producing said antibiotic directly from fortimicin A in a four step process which results in a 50-60 percent yield of product.

Method of producing 2-deoxyfortimicin A

Described are novel derivatives of seldomycin factor 5 (XK-88-5) and particularly 3'-Epi-seldomycin factor 5, which exhibit improved activity against gram-positive and gram-negative bacteria resistant to aminoglycoside antibiotics, and a method of preparing them.

Seldomycin factor 5 derivatives

Described are novel derivatives of seldomycin factor 5 (XK-88-5) and particularly 3'-deoxyseldomycin factor 5, which exhibit improved activity against gram-positive and gram-negative bacteria resistant to aminoglycoside antibiotics, and a method of preparing them.

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Gurnee, IL, United States of America

Ronald E Carney