Search Images Maps Play YouTube News Gmail Drive More »
Sign in
Screen reader users: click this link for accessible mode. Accessible mode has the same essential features but works better with your reader.


  1. Advanced Patent Search
Publication numberUS2920012 A
Publication typeGrant
Publication dateJan 5, 1960
Filing dateSep 1, 1955
Priority dateSep 1, 1955
Publication numberUS 2920012 A, US 2920012A, US-A-2920012, US2920012 A, US2920012A
InventorsRobert G Sanders, Roland R Read, Richard J Palazzolo
Original AssigneeWarner Lambert Pharmaceutical
Export CitationBiBTeX, EndNote, RefMan
External Links: USPTO, USPTO Assignment, Espacenet
Therapeutic compositions for inhibiting carbonic anhydrase activity
US 2920012 A
Abstract  available in
Previous page
Next page
Claims  available in
Description  (OCR text may contain errors)

United States Patent "face 2,920,012 Patented Jan. 5, 1960 Robert G. Sanders, Clayton, Roland R. Read, Ladue, and Richard J. Palaz'zolo, Alfton, Mo., assignors to Warner-Lambert Pharmaceutical Company, St. Louis, Mo., a corporation of Delaware No Drawing. Application September 1, 1955 Serial No. 532,113

7 Claims. (Cl. 167-515) This invention relates to therapeutic compositions and 2 more particularly to therapeutic compositions which are eifective to inhibit carbonic anhydrase activity.

This application is a continuation-in-part of our applik cation Serial No. 456,639, filed September 16, ,1954, now -abandoned.

Briefly, the present invention is directed to therapeutic compositions comprising a compound having the fortmula:

NHCOOR -wherein Ris-ethyl, p-hydroxy ethyl or allyl, auda' pharmaceutical carrier. The invention also includes the method of inhibiting carbonic anhydrase activity through the use of the above-noted compounds.

Among the several objectsof the invention may'be noted the provision of therapeutic compositions and methods which are efiective to inhibit the activity of the enzyme carbonic anhydrase; the provision of such therapeutic compositions which are effective diuretic compositions; and the provision of therapeutic compositions of the class described which are stable and nontoxic. Other objects and features will be in part apparent and in part pointed out hereinafter. a

- The invention accordingly comprises the products and methods hereinafter described, the scope of the invention being indicated in the following claims.

The enzyme carbonic anhydrase plays an important role in the acidification of the urine through its catalysis of the reversible reaction:

The carbonic acid so formed yields hydrogen and bicarbonate ions through the reversible reaction:

'Thus, carbonic anhydrase activity indirectly determines the number of hydrogen ions which are available to re- .place sodium ions inbuffercompounds from the blood, suchas, for example, disodium phosphate-rand sodium bicarbonate. The sodium .ions thus replaced are conserved and return to the blood, and carbonic acid formed in the above reaction catalyzed by carbonic anhydrase also returns to the blood.

In accordance with the present invention, it has been found that carbonic anhydraseactivity can be effectively inhibited and the excretiontof water, sodium and potassium in the urine increased by the use of therapeutic compositions including a p-sulfamyl carbanilate compound having the following formula:

SiOgNHg NHCOOR wherein R is ethyl, S-hydroxy ethyl or allyl. The inhibition of carbonic anhydrase activity by the composi tionsof the present invention resultsin a reduction of hydrogen ions available for the acidification of the urine. As a consequence, the pH of the urine increases, fewer sodium ions of the buffer compounds are replaced by hydrogen ions, and larger amounts of sodium, chloride and bicarbonate are excreted in the urine. Further, because the formation of carbonic acid from carbon dioxide and water is inhibited, water excretion is increased. Thus, the diuretic action provided by the compositions of the inventionis useful to relieve edema, for example, caused bycongestive heart failure or other causes.

It has further been found in accordance with the present invention that when 2,4-diamino s-triazine (also known as formoguanamine), 2-arnino-4-B-carboxy ethyl amino, s triazine, 2,4-(fl-carboxy ethyl amino) s-triazine, 2 amino 4 anilino 1,3,5 triazine, 2 amino 4- (p-chloroanilino)-l,3,5-triazine or the nontoxic salts of one of these compounds is included in these novel therapeutic compositions, the amount of water, sodium and chloride, particularly the latter, secreted in the urine is markedly increased. The diuretic action of these triazine compounds is believed to be due to the inhibition of tubular reabsorption brought about by a partial anoxia from the inhibition of respiratory enzymes involved in the tubular reabsorption of water and sodium chloride. The increase in diuretic action noted when one of these'tria'zine compounds is included in the therapeutic" compositions'of the invention is more than can be accounted for by the additive eifect of the carbanilate compound andthe triazine compound on the basis of their action individually, as' is shown by the tests described hereinafter. Among the exemplary nontoxic salts of 2, 4- diamino s-triazine, 2-amino-4-[3-carboxy ethyl amino s-triazine, 2,4-(f3-carboxy ethyl amino) striazine, 2- amino-4-ani1ino-1,3,5-triazine and 2-amino-4-(p-chloroanilino)-l,3,5-triazine which are useful as components of the compositions of the invention may be mentioned the hydrochloride, sulfate, phosphate and tartrate salts. The salts of the alkali metals, ammonia and amines may also be utilized. The amount of triazine component, when includedin the compositions of the invention, may be varied considerably and may be, for example, approximately 27% by weight.

The p-sulfamyl carbanilate compounds specified herein are not only compatible with other components employed in therapeutic compositions, but they are also stable and nontoxic. For example, intraperitoneal doses of; p-Sulfamyl fl-hydroxy ethyl carbanilate .up to 1600 mg/kg, were tolerated by. male white rats, and only three animalsout of .a group of ..nine. receiving 1800 Ing /kg. intraperitoneally died.1.Moreover,. oral doses up to 1500 rngjlgg. of this carbanilate. compound did not prpduce any ill effectjn male whiterats. Three groups of white rats, with six animals in each group, received 6 00 QOQ and 1200 mg./kg. respectively of p-sulfamyl fl-hydr'oxy ethyl carbanilate-per day for twelve days and gross pathology after being sacrificed showed "a" normal and autopsied on the thirteenth day.

The carbanilate compounds of the novel therapeutic compositions of the invention have the formula:


wherein R represents ethyl, B-hydroxy ethyl or allyl. The therapeutic compositions of the present invention comprise a compound of the above mentioned class and a pharmaceutical carrier which may be either a liquid or solid material. It is preferred that the pharmaceutical carrier be a solid material such as, for example, starch, gelatin, lactose, talc, calcium stearate or the like. Any of the pharmaceutical carriers conventionally used in therapeutic compositions may be utilized where such materials are compatible with the aforementioned carbanilate compounds. These compositions include tablets, capsules and the like, and may include a liquid pharmaceutical carrier and be in the form of solutions, dispersions, suspensions, elixirs and the like.

The percentage of the carbanilate compound incorporated in the therapeutic compositions of the invention may be varied considerably. Exemplary compositions may contain approximately 80% by weight of one of the above mentioned carbanilate compounds, although greater or smaller amounts than 80% may also be used in the practice of the invention. The daily dosage administered to a patient may also vary considerably, depending upon the patients condition and the amount prescribed by the physician in charge. In the case of edema, the daily dosage of these novel compositions may be such that the patient ingests on the order of 250 mg. to 500 mg. of one of the carbanilate compounds per day.

The following examples illustrate the invention.

Example 1 Therapeutic diuretic tablets were prepared having the following composition:

Component: G. p-Sulfamyl fi-hydroxy ethyl carbanilate 0.500 Formoguanamine 0.250 Betaine hydrochloride 0.065 Corn starch U.S.P. 0.100 Calcium stearate 0.010 Talcum powder 0.005

Example 2 Therapeutic diuretic tablets were prepared having the following composition:

Component: G. p-Sulfamyl S-hydroxy ethyl carbanilate 0.250

2 amino 4 anilino-1,3,5-triazine hydrochloride 0.125 Betaine hydrochloride 0.065 Corn starch U.S.P. 0.100 Calcium stearate 0.010 Talcum powder 0.005

The tablets were prepared by first mixing the p-sulfamyl fl-hydroxy ethyl carbanilate, 2-amino-4-anilino-l,3,5-trtazine hydrochloride and betaine hydrochloride in the dry state. This mixture was then wet granulated with isopropyl alcohol (0.125 ml. per tablet) and water (0.125 ml. per tablet). The wet granulated mixture was thereafter dried on trays at a temperature of F. for 15 hours, and the dry mixture was ground in an oscillator using a No. 11 heavy wire screen. The corn starch, calcium stearate and talcum powder were added and, after thorough mixing, the mixture was compressed into tab ets.

Example 3 The inhibition of carbonic anhydrase activity by psulfamyl allyl carbanilate was determined as follows:

The carbonic anhydrase employed was prepared from rabbit red blood cells by the method of Keilin and Mann (Biochem. J. 34, 1166, 1940). The red blood cells were Washed three times with saline by centrifugation. To the washed cells (10 ml.) in a centrifuge tube were added distilled water (6 ml.), ethyl alcohol (4 ml.) and chloroform (5 ml.). The mixture was shaken well and centrifuged for 25 minutes at 2500 r.p.m. A three phase system was formed consisting of a top layer of enzyme solution, a central layer of denatured protein and a bottom layer of chloroform.

The top layer was removed and the activity of the carbonic anhydrase determined by a modification of the meth 0d of Meldrum and Roughton (J. Physiol. 80, 116-124, 1934), which measures the catalytic effect of the carbonic anhydrase enzyme on the rate of evolution of carbon dioxide from sodium bicarbonate solutions when mixed with phosphate buffer of pH 6.8. A sodium bicarbonate solution (1.1 ml.) (0.2 M) (prepared by dissolving the sodium bicarbonate in a 0.02 M sodium hydroxide solution) was placed in the main compartment of a 15 ml. Warburg flask while phosphate buffer (1 ml.) made by mixing equal volumes of disodium phosphate solution (0.2 M) and potassium acid phosphate solution (0.2 M) was placed in one side arm of the flask. The flask was attached to a manometer and the solution equilibrated 1 at 25 C in a shaking machine. The rate of carbon dioxide evolution in the presence of carbonic anhydrase enzyme solution (1 ml.) was measured and compared with a blank. It was found that at a concentration of l mg./ml. of p-sulfamyl allyl carbanilate 84% inhibition of carbonic anhydrase activity was effected when tested according to this method.

Other compositions of the invention corresponding to the above described compositions andgcontaining comparable amounts of p-sulfamyl ethyl carbanilate, p-sulfamyl, allyl carbanilate and one of the aforementioned triazine compounds may be prepared and are effective in the practice of the invention.

It will be understood, of course, that the therapeutic compositions of the invention may be employed in the treatment of any condition where the inhibition of carbonic anhydrase activity will produce a beneficial result.

In view of the above, it will be seen that the several objects of the invention are achieved and other advantageous results attained.

As various changes could be made in the above products and methods without departing from the scope of the invention, it is intended that all matter contained in the above description shall be interpreted as illustrative and not in a limiting sense.

We claim:

1. A therapeutic composition comprising a compound having the formula:

NHCOOR wherein R represents a radical selected from the group consisting of ethyl, p-hydroxy ethyl and allyl, a compound selected from the group consisting of 2,4-diamino s-triazine, 2-amiuo-4-p-carboxy ethyl amino s-triazine, 2,4- (B-carboxy ethyl amino) s-triazine, 2-amino-4-anilino- 1,3,5-triazine, 2-amino-4-(p-chloroa-nilino)-1,3,5-triazine and the nontoxic salts thereof, and a pharmaceutical carrier.

2. A therapeutic composition comprising p-sulfamyl a hydroxy ethyl carbanilate, 2-amino-4- -anilino-1,3,5-triazine and a pharmaceutical carrier.

3. A therapeutic composition comprising p-sulfamyl ethyl carbanilate, 2-amino-4-anilino-1,3,5-triazine and a pharmaceutical carrier.

4. A therapeutic composition comprising p-sulfamyl allyl carbanilate, 2-amino-4-anilino-1,3,5-triazine and a pharmaceutical carrier.

5. A therapeutic composition comprising p-sulfamyl References Cited in the file of this patent U.S. Dispensatory, 25th ed., pp. 191-492.

J. Am. Chem. Soc., vol. 72, November 1950, pp. 4893-4896.

Chem. Abst., vol. 41, 1947, pp. 4809-4811.

Vogl: Diuretic Ther., Williams and Wilkins Co., 1953, p. 47, Baltimore, Md.

I.A.M.A., vol. 151, No. 16, Apr. 18, 1953, p. 1430.

Non-Patent Citations
1 *None
Referenced by
Citing PatentFiling datePublication dateApplicantTitle
US4917108 *Jun 29, 1988Apr 17, 1990Mault James ROxygen consumption meter
US5836300 *Mar 11, 1997Nov 17, 1998Mault; James R.Metabolic gas exchange and noninvasive cardiac output monitor
US6135107 *Nov 13, 1998Oct 24, 2000Mault; James R.Metabolic gas exchange and noninvasive cardiac output monitor
US6277645Sep 3, 1999Aug 21, 2001James R. MaultMethod and apparatus for respiratory gas analysis employing measurement of expired gas mass
US6309360Jan 16, 1998Oct 30, 2001James R. MaultRespiratory calorimeter
US6402698Feb 5, 1999Jun 11, 2002James R. MaultMetabolic calorimeter employing respiratory gas analysis
US6406435Nov 17, 1999Jun 18, 2002James R. MaultMethod and apparatus for the non-invasive determination of cardiac output
US6468222Aug 2, 2000Oct 22, 2002Healthetech, Inc.Metabolic calorimeter employing respiratory gas analysis
US6478736Oct 10, 2000Nov 12, 2002Healthetech, Inc.Integrated calorie management system
US6482158May 14, 2001Nov 19, 2002Healthetech, Inc.System and method of ultrasonic mammography
US6506608Aug 20, 2001Jan 14, 2003Healthetech, Inc.Method and apparatus for respiratory gas analysis employing measurement of expired gas mass
US6517496May 10, 2000Feb 11, 2003Healthetech, Inc.Airway-based cardiac output monitor and methods for using same
US6607387Oct 26, 2001Aug 19, 2003Healthetech, Inc.Sensor system for diagnosing dental conditions
US6612306Oct 11, 2000Sep 2, 2003Healthetech, Inc.Respiratory nitric oxide meter
US6620106Oct 1, 2001Sep 16, 2003Healthetech, Inc.Indirect calorimetry system
US6629934Feb 1, 2001Oct 7, 2003Healthetech, Inc.Indirect calorimeter for medical applications
US6645158Apr 23, 2002Nov 11, 2003Healthetech, Inc.Metabolic calorimeter employing respiratory gas analysis
US6790178 *Sep 25, 2000Sep 14, 2004Healthetech, Inc.Physiological monitor and associated computation, display and communication unit
US6899683May 31, 2002May 31, 2005Healthetech, Inc.Metabolic calorimeter employing respiratory gas analysis
US6955650May 31, 2002Oct 18, 2005Healthetech, Inc.Metabolic calorimeter employing respiratory gas analysis
US7291114Apr 1, 2003Nov 6, 2007Microlife CorporationSystem and method of determining an individualized drug administration protocol
US7392193Jun 18, 2001Jun 24, 2008Microlife CorporationSpeech recognition capability for a personal digital assistant
US20010044588 *Mar 29, 2001Nov 22, 2001Mault James R.Monitoring system
US20020047867 *Sep 7, 2001Apr 25, 2002Mault James RImage based diet logging
US20020055857 *Oct 25, 2001May 9, 2002Mault James R.Method of assisting individuals in lifestyle control programs conducive to good health
US20020133378 *Oct 12, 2001Sep 19, 2002Mault James R.System and method of integrated calorie management
US20020138213 *Mar 4, 2002Sep 26, 2002Mault James R.System and method of metabolic rate measurement
US20030023181 *Jul 25, 2002Jan 30, 2003Mault James R.Gas analyzer of the fluorescent-film type particularly useful for respiratory analysis
US20030065274 *Jun 10, 2002Apr 3, 2003Mault James R.Method of respiratory gas analysis using a metabolic calorimeter
US20030065275 *Oct 16, 2002Apr 3, 2003Mault James R.Metabolic calorimeter employing respiratory gas analysis
US20030105407 *Oct 8, 2002Jun 5, 2003Pearce, Edwin M.Disposable flow tube for respiratory gas analysis
US20030130567 *Jan 8, 2003Jul 10, 2003Mault James R.Health-related devices and methods
US20030130595 *Jan 10, 2003Jul 10, 2003Mault James R.Health improvement systems and methods
US20030152607 *Sep 27, 2002Aug 14, 2003Mault James R.Caloric management system and method with voice recognition
US20030163321 *Jun 18, 2001Aug 28, 2003Mault James RSpeech recognition capability for a personal digital assistant
US20030167016 *Feb 10, 2003Sep 4, 2003Mault James R.Airway-based cardiac output monitor and methods for using same
US20030208110 *May 25, 2001Nov 6, 2003Mault James RPhysiological monitoring using wrist-mounted device
US20030208133 *Jun 6, 2001Nov 6, 2003Mault James RBreath ketone analyzer
US20030220579 *Apr 1, 2003Nov 27, 2003Mault James R.System and method of determining an individualized drug administration protocol
US20030226695 *May 24, 2001Dec 11, 2003Mault James R.Weight control method using physical activity based parameters
USD478660Jul 1, 2002Aug 19, 2003Healthetech, Inc.Disposable mask with sanitation insert for a respiratory analyzer
U.S. Classification514/155, 514/869, 560/13, 544/205, 514/245, 544/206
International ClassificationA61K31/195
Cooperative ClassificationA61K31/195, Y10S514/869
European ClassificationA61K31/195