KR20040081135A - Method of treatment of gastrointestinal disease and polymeric composition for use therein - Google Patents

Abstract

The present invention relates to polymers comprising derivatives of poly (2-propene, 2-propene acid) formed by reacting alcohol or phenol with poly (2-propene, 2-propene acid) to form a protected carbonyl group. A method of treating gastrointestinal disorders comprising administering an antimicrobial agent. The present invention also relates to a composition for treating gastrointestinal diseases.

Description

METHOD OF TREATMENT OF GASTROINTESTINAL DISEASE AND POLYMERIC COMPOSITION FOR USE THEREIN}

Antimicrobials are compounds that kill microorganisms such as bacteria. Antibiotics are a type of antimicrobial agent usually derived from other microorganisms and act by interfering with certain mechanisms in the target microorganism. Antibiotics were used for the first time between the 1940s and 1950s, and their use has increased ever since. However, the development of antibiotic resistance has had serious and serious consequences that are life threatening worldwide. Some strains of Staphylococcus are resistant to almost all antibiotics and cause fatal infections in hospitals. Other drug resistant organisms include pneumococci, which cause pneumonia, and cryptosporidium and E. coli, which cause diarrhea.

The use of antibiotics in animal feed is largely believed to be the cause of the accelerated development of resistance, and consequently many countries regulate its use. This causes the problem of animal breeding, making it difficult to suppress disease and obtain optimum growth rates. This was especially a problem when raising pigs and poultry. For example, gastrointestinal disorders such as E. coli in pigs and Coccidiosis in poultry can have overwhelming effects.

Melrose et al., In International Patent Application Publication No. 96/38186, disclose for the first time a method for preparing acrolein polymer for use in the treatment of gastrointestinal diseases.

This polymer has a repeating unit represented by the formula (I) or this unit in hydrated form, hemiacetal form or acetal form represented by the following formulas (a) to (f) has been demonstrated previously:

Wherein R is hydrogen and n is an integer of 1 or more. Prior to this polymer, the bactericidal properties of acrolein polymers as preservative applications have been described by Melrose et al. In International Application No. WO88 / 04671. German patent application P4404404 and corresponding applications, such as EP667358 and AU 11686/95 (currently expired), disclose a process for the polymerization of acrolein in aqueous sodium hydroxide solution. The specification states that the final product, polyacrolein, is dissolved in the polyhydric alcohol at 40-50 ° C. to form a polyacrolein solution in the polyhydric alcohol. As described below, the assignee of this German patent application subsequently discovered the problem of the polymer and showed low solubility in aqueous media.

In European Publication No. 792895 Walley et al. (Corresponding to US 6060571) disclose acrolein releasing polymers prepared by copolymerization of acrolein monomers and polyhydric alcohols. Wally et al. Found that the polyacrolein described in German application No. P4404404 was problematic in that it was lower than the desired yield and was virtually insoluble in water. European application 792895 teaches to solve this problem by copolymerizing acrolein monomers with polyhydric alcohol monomers to form acrolein releasing polymers. The proposed structure of the copolymer is as follows:

[expression]

Free acrolein acts as an antimicrobial agent but is irritating to the eyes, lungs, tissues and skin. Thus, there is a need for an antimicrobial agent that is stable in a wide range of industrial applications, particularly gastrointestinal treatments, and is highly water soluble and safe to use. There is also a need for an antibacterial agent that is effective in the treatment or prevention of gastrointestinal diseases that can reduce the pressure of antibiotic resistant development.

The above discussion of the background of the present invention is intended to explain the research background of the present invention. Accordingly, it should not be taken as an endorsement that all of the materials described are publicly known or known or were part of common sense on the priority date of all claims.

The present invention relates to methods of treating or preventing gastrointestinal diseases, methods of promoting growth of animals, and antimicrobial compositions for use in such treatments.

The inventors have found that the poly (2-propenal, 2-propenoic acid) polymers used for the treatment or prevention of gastrointestinal diseases can be reacted with alcohols or polyols to form protected carbonyl groups such as acetal and / or hemiacetal derivatives. And stability have been found to increase significantly. Surprisingly, the inventors have found a substantial increase in the activity of the derivatives in the treatment or prevention of gastrointestinal diseases, despite the extremely low or almost negligible amount of free acrolein in this polymer. In addition, the polymer was also very water soluble.

The present invention relates to an organic compound containing at least one hydroxyl group, such as an alcohol selected from alkanols, phenols, polyols and mixtures thereof, in order to form protected carbonyl groups. A method of treating or preventing gastrointestinal diseases in an animal (including humans) is provided, including administering to the animal an effective amount of a poly (2-propeneal, 2-propene acid) derivative formed by reaction with the same.

In another aspect, the present invention relates to an organic compound containing at least one hydroxyl group, such as an alcohol selected from alkanols, phenols, polyols and mixtures thereof, for forming protected carbonyl groups. Provided is an antimicrobial agent for treating gastrointestinal diseases, comprising a poly (2-propenal, 2-propene acid) derivative formed by reacting with 2-propene acid).

The term polyol, as used herein, means a molecule containing at least two hydroxyl groups.

The derivatives formed are generally selected from hemiacetal derivatives and acetal derivatives. While not wishing to be bound by the following theory, the reaction of poly (2-propeneal, 2-propenoic acid) with alcohols forms hemiacetal groups and / or acetal groups from at least some of the aldehyde side groups to form carbonyl groups of the polymer It is believed to have a stabilizing effect against alkali decomposition by (Cannizzaro) reaction. Formation of acetal groups has been observed to significantly reduce or eliminate the release of free acrolein, but surprisingly increases the activity of the final product derivative.

In another embodiment, the present invention provides the use of the aforementioned antimicrobial agent, which is useful for the manufacture of a medicament for the treatment or prevention of gastrointestinal diseases.

The terms "comprises" and variations thereof, such as "comprising" and the like, as used throughout the description and claims of this specification are not intended to exclude other additives or other components or integers.

The antimicrobial agents of the present invention can be prepared by heating poly (2-propaneal, 2-propenoic acid) in the presence of an alcohol, preferably a polyol such as polyethylene glycol. Water is always present in alcohols, and it is thought that the presence of at least some water helps the nucleophilic reaction to form hemiacetal or acetal.

The heating of the solution is generally carried out at a temperature in the range from 40 to 150 ° C., more preferably in the range from 40 to 115 ° C. and most preferably in the range from 70 to 115 ° C.

The antimicrobial agents of the present invention are prepared from poly (2-propenal, 2-propene acid) polymers. This polymer and its preparation are described in International Patent Publication No. WO96 / 38186 (PCT / AU96 / 00328), which is incorporated herein by reference. The poly (2-propeneal, 2-propenoic acid) polymer is preferably prepared by polymerizing acrolein, preferably by anionic polymerization, in aqueous solution, followed by automatic oxidation. The polymers thus prepared comprise repeating units of formula (I) and at least one hydrated form, hemiacetal form and acetal form (generally in mixtures).

As such, the hydrated, hemiacetal and acetal forms produced by the polymerization of acrolein are known to occur by various carbon-carbon and carbon-oxygen polymerization mechanisms of acrolein. For example, the hydrated form is generally a hydrated diol form, the hemiacetal or acetal form can be prepared by condensing the diol form with an aldehyde or diol form, and the tetrahydropyran form or fused tetrahydropyran form It can be prepared by condensing the diol form and the aldol-michael autocondensation form. General examples of this form are as shown in the following formulas (a) to (f):

[Formula I]

[Formula a]

[Formula b]

[Formula c]

[Formula d]

[Formula e]

[Formula f]

Wherein R is hydrogen and n is an integer of 1 or more. The proportion of repeat units in formula (I) is generally less than 20%, more generally in the range of 5 to 15%. Although the ratio of these units is relatively low, we have found that these units have a significant effect on the stability of the polymer.

Poly (2-propeneal, 2-propenoic acid) may generally contain up to 10% of monomers other than acrolein on a molar basis of monomer units, but most preferred is acrolein homopolymer (pre-autooxidation). Other monomers may be selected from the group consisting of acrylic acid and vinyl pyrrolidone if used. 2-propene acid groups are generally present in amounts of 0.1 to 5 mol / kg of carboxyl groups. Poly (2-propeneal, 2-propene acid) polymers generally have a number average molecular weight of at least 1000, most preferably at least 2000. Generally, the molecular weight is less than 10,000.

The antimicrobial agents of the present invention are derivatives of poly (2-propenal, 2-propenoic acid) prepared by reaction with alcohols or phenols to form protected carbonyl groups. Protected carbonyl groups are formed from 2-propenal groups, which react with alcohol to form hemiacetal groups and acetal groups. The alcohol is preferably a polyol which means containing at least two hydroxyl groups. Alkanols such as C ₁ to C ₁₀ may also be used. If the alcohol is a polyol, this reaction may produce acetal or hemiacetal formed by the reaction of one or more alcohol groups.

With respect to the above formula (I), hemiacetal form and acetal form, the present invention relates to derivatives having a lower number of units of formula (I) in which one or more R groups are derived from alcohols or two or more R groups together when the alcohol is a polyol To yield derivatives capable of forming crosslinking groups such as cyclic acetal groups.

The likelihood that polyols will form with internal cyclic groups will depend on the spacing and arrangement of the polyols. Preferred alcohols are polyalkylene glycols and more preferred alcohols are polyethylene glycols.

The molecular weight of the polyalkylene glycol is preferably in the range from 200 to 2000, more preferably in the range from 200 to 1000.

During the preparation of the antimicrobial polymer, alcohols such as polyethylene glycol are preferably present in an amount between 50 and 99% by weight. Relatively dilute acrolein polymer compositions are particularly preferred when the alcohol is a polyol because the incidence of intermolecular crosslinking decreases upon dilution.

More preferably, polyethylene glycol is present in an amount between 64 and 95% by weight in the preparation of the polymer.

Subsequently, it is preferred to add a base or an alkali to the polymer, and then abruptly change it to an acidic pH before and / or with heating, since the antibacterial activity of the polymer is enhanced by neutralizing the acid groups of the polymer. The addition of base or alkali is preferred to first bring the pH of the poly (2-propeneal, 2-propene acid) polymer to between 7 and 9. Even more preferably, the initial pH at the time of base addition is about 8. The base is preferably alkali metal hydroxide, carbonate, bicarbonate or mixtures thereof.

In another form of the invention, the release of the free acrolein monomer is inhibited from continuous release, thereby making the polymer less likely to provide a source of tissue or skin irritation.

The inventors have found that the antimicrobial agent of the present invention has a significantly improved activity in the inhibition of gastrointestinal diseases when compared to its poly (2-propeneal, 2-propene acid). The overactivated derivatives of the present invention can be used to treat various animals (including humans) and various microbial infections.

The antimicrobial agents of the present invention can be used to treat gastrointestinal diseases in humans, but the treatment of other animals, particularly animals selected from the group consisting of dogs, pigs, sheep, horses, goats, cattle, cats, poultry, ducks, turkeys and quails. It is especially preferable because it can be used for.

The antimicrobial agent of the present invention can be formulated for oral or rectal administration. Rectal administration may be particularly useful for ruminants. In addition, oral formulations of ruminants can be prepared using enteric coating to provide optimal activity in the latter half of the gastrointestinal tract.

The antimicrobial agents of the present invention are particularly useful for the treatment and prevention of gastric ulcers, diarrhea and gastrointestinal cancers. The antimicrobial agents of the present invention can also be used to improve the rate of conversion of animal feed to body weight, thereby improving the rate of weight gain of the animal.

The inventors have found that the antimicrobial agents of the present invention can be used as growth promoters and that the polymers of the present invention can be used in place of the currently used antibiotics. Drug resistance of pathogenic bacteria is a major clinical problem in human drugs. This problem is magnified when major antibiotics are used in animal feed to gain weight in livestock animals, especially poultry and pigs. In fact, the use of conventional antibiotics in animal feed is prohibited in some European countries. The antimicrobial agents of the present invention can be used in the treatment of animals to significantly extend the shelf life of antibiotics conventionally used in human treatment.

We have found that the antimicrobial agents of the invention are effective against a variety of microorganisms, including protozoa, Gram-positive bacteria and Gram-negative bacteria. The polymers of the present invention may contain multiple structures of various arrangements, so that they can be found to be optimal for proteins found in the cell walls of target organisms. Accordingly, the protein inactivation and cell destruction rate can be increased. The antimicrobial agents of the present invention have been found to be particularly useful for providing a wide range of activities against E. coli or enterobacteriaceae among Gram-negative bacteria. In particular, it is useful for the treatment of gastrointestinal diseases caused by E. coli infections such as enterotoxin E. coli and β-hemolytic E. coli. Escherichia coli is a threat to the swine business. The disease is generally associated with the proliferation of β-hemolytic E. coli in the small intestine of post-weaning pigs and causes high mortality and incidence in young weaning piglets. Infected young pigs will not gain weight normally.

Escherichia coli is a protozoan disease of animals, especially poultry, which, if left untreated, has a threatening effect. We have found that the antimicrobial agents of the invention can be used to treat or prevent E. coli in birds, especially poultry. Clinical signs of common E. coli in chicks include vigorous growth reduction, rapid weight loss, diarrhea and dysentery. The most serious effects are found in the intestine, where the protozoa invades the mucosa, causing epithelial damage, lesions and bleeding. Vaccines have been used as an attempt to prevent E. coli, but this has had side effects, including a tendency to reduce body weight and breeding efficiency.

The antimicrobial agents of the present invention can be used with other drugs known to be active against E. coli. Such drugs include nitro-carbanilides, quinoline, pyridones, guanidines, quinoxaline, tortralral, toluamides, fortified sulfa drugs, and ion penetrating carriers with carbanilides.

Clostridia are Gram-positive bacteria that cause serious diseases in a variety of animals. For example, necrotic enteritis is a disease known to affect commercial poultry. Clostridia bacteria produce exotoxins, some of which are the most poisonous of all known toxins. Necrotic enteritis affects the spirit world, especially from 14 to 42 days of age. This disease can lead to apparent numb diarrhea, leading to death within hours.

More serious gastrointestinal diseases, such as chronic gastritis, gastric ulcer and duodenal ulcer, are serious health problems in humans. Helicobacter is believed to be responsible for the development of ulcers and gastrointestinal cancers, especially adenocarcinoma of the stomach. The inventors have found that the antimicrobial agents of the invention are particularly useful against Helicobacter bacteria, including Helicobacter pylori, in gastrointestinal diseases in animals, especially humans.

Stomach infection by Helicobacter pylori is one of the most common infectious diseases worldwide. About 50% of the population is infected with H. pylori. In developing countries, it is estimated that over 80% of the population is already infected with H. pylori during childhood.

Helicobacter pylori is a gram negative, aerobic, helical bacterium that is mobile by flagella at one end of a cell. The basic treatment for H. pylori infection is the so-called three-way antibiotic treatment, which includes either metronidazole or clarithromycin. Unfortunately, strains of H. pylori have been found to be resistant to all of these antibiotics.

H. pylori grows in the gap between the epithelial cell surface of the stomach and the layer of mucosal gel layered on it. H. pylori is also found on the surface of gastric epithelial cells in the duodenum and esophagus. Other animal species have their own unique Helicobacter species in their gastrointestinal tract that have properties similar to H. pylori. In addition to gastric cancer, H. pylori is directly related to gastritis and peptic ulcer formation in humans.

In general, Helicobacter spp., Specifically H. pylori, urea, which hydrolyze urea to secrete ureases that generate ammonia and bicarbonate ions, thereby increasing the pH of the environment adjacent to the bacteria and thus survives the above extreme conditions. This change in local conditions protects the bacteria from the bactericidal effects of gastric acid. In addition, the position of the bottom of the protective mucosal layer above is advantageous for survival and its mobility allows it to reach this position through the mucosal layer.

Epithelial cells inside the stomach are inherently difficult to penetrate, which is part of the epithelial function that protects the rest of the body from gastric acid and gastric juice. This difficulty of penetration also makes it difficult for the body's natural defenses to cross the stomach wall and reach the H. pylori infection site. This results in two outcomes, one in which the body supplies more nutrients to the site of infection to support white blood cells, T cells and other defense mechanisms, while at the same time supplying bacteria; The other is that the protective cells actually die, releasing their superoxide ions and other lethal compounds, damaging surrounding epithelial cells.

Obviously, this activity causes gastritis, which can easily progress to peptic ulcers. If this damage persists, the likelihood of developing gastric adenocarcinoma and mucosal associated lymphoid tissue (MALT) lymphoma increases significantly. Adenocarcinoma of the stomach begins in the mucosa, and intestinal metaplasia, the first stage of the onset, is a gastric reaction to remove H. pylori infection by itself. In addition, a study conducted at the UCL Medical Institute found that MALT lymphomas require the help of H. pylori specific T cells to proliferate. Thus, the treatment of H. pylori infection was found to be very effective in treating MALT lymphoma. The World Health Organization has classified this pathogen as a Class I carcinogen.

Accordingly, the present invention is made by reacting poly (2-propenal, 2-propenic acid) with an organic compound containing a hydroxyl group selected from alkanols, phenols, polyols and mixtures thereof to form protected carbonyl groups. Provided are methods for treating or preventing diseases of the gastrointestinal tract caused by Helicobacter infection, including gastrointestinal administration of a therapeutically effective amount of a formulation comprising a poly (2-propeneal, 2-propene acid) derivative. The term polyol, as used herein, means a compound containing at least two hydroxyl groups. The derivative formed is generally one selected from hemiacetal derivatives and acetal derivatives.

Thus, the methods of the present invention may be used in place of chemotherapy or radiation therapy or surgery that is customary for the treatment of gastrointestinal cancer.

The present invention also relates to poly (2-propenal, 2-, formed by reacting an organic compound containing at least one hydroxyl group with poly (2-propenal, 2-propenic acid) to form a protected carbonyl group. Provided are methods of treating gastrointestinal infections caused by Helicobacter fungus, such as gastritis, gastric ulcer, duodenal ulcer, gastric malignant lymphoma or gastric cancer, including gastrointestinal administration of a therapeutically effective amount of a formulation comprising a propene acid) derivative.

The present invention provides an alternative to the general treatment of Helicobacter infection, including the so-called three-way antibiotic therapy, which generally comprises either metronidazole or clarithromycin. H. pylori has been shown to be resistant to both of these antibiotics, and the inventors have found that the method of the present invention can effectively treat such antibiotic resistant bacteria.

Formulations of the present invention which are reaction products between an organic compound containing one or more hydroxyl groups and a poly (2-propeneal, 2-propenoic acid) are equivalent to a non-overactive poly (2-propeneal, 2- More effective in treating Helicobacter infection than propene acid).

The present invention also provides the use of a poly (2-propenal, 2-propenoic acid) derivative used in the manufacture of a medicament for the treatment or prevention of a disease caused by a Helicobacter infection.

The methods of the present invention can be used to treat or prevent gastrointestinal cancer. These gastrointestinal cancers include, for example, esophageal cancer, gastric cancer, intestinal cancer, and colon cancer. One example of this type of cancer is the human colon cancer cell line HT-29.

When the antimicrobial agent of the present invention is mixed with animal feed or water, it can be mixed and used in a conventional manner. In a preferred embodiment, the antimicrobial agents of the present invention may be mixed in the form of premixes. This premix preferably comprises an antimicrobial agent, a physiologically acceptable carrier and optionally a feed. This premix is generally in a relatively concentrated form, which can be diluted into feed with other ingredients, such as one or more other carriers, vitamins and mineral supplements, to produce the final feed of the animal. This premix preferably comprises an antimicrobial agent at a concentration in the range of 0.1 to 70% by weight, more preferably in the range of 0.5 to 50% by weight. The optimal concentration depends on whether the treatment is prophylactic, inhibitory or therapeutic and whether the antimicrobial agent of the present invention is the only active agent or used in the simultaneous treatment with other materials or antimicrobial agents.

In a preferred embodiment, the concentrated composition of the antimicrobial agent is preferably controlled release. Controlled release forms include antimicrobials and polymeric materials for controlled release of the antimicrobials from controlled release systems, and are particularly useful in compositions for addition to solid feeds. Controlled release formulations may consequently delay the release of the antimicrobial agent so that release occurs primarily in the duodenum. Controlled release polymers can also be used as rectal suppositories or to minimize rejection of the composition due to taste.

The antimicrobial composition according to the invention may be in the form of pills, pills or similar solid compositions. Pills containing the antimicrobial agent of the present invention may be prepared as follows:

(i) dissolving the antimicrobial agent in an aqueous alkali or base solution;

(ii) neutralizing this solution with acid;

(iii) adding an insoluble crosslinked absorbent acrylic acid polymer and / or a copolymer of acrylamide and acrylic acid to the neutralized solution to form wettable swollen pills; And

(iv) optionally, completely or partially drying the wettable swollen pill.

The wettable swollen pills thus prepared can be used, for example, in the wet, partially dried or completely dried state as an additive to animal feed. The system is also designed so that the polymer in question can be substantially contained in the matrix by the carboxyl-containing groups on the outer surface of the polymer matrix even when in the acidic environment of the stomach. However, in the alkaline environment of the duodenum, the carboxyl groups of the polymer matrix are ionized to repel each other, and the pills swell rapidly, causing repulsion between their ionic groups to speed up the passage of the polymer through the duodenum. It is excluded through the diffusion process.

In the present invention, the term "controlled release system" includes examples of the range as recited in the document "Controlled Drug Delivery" (Robinson & Lee, 1987) and is used in the same context. Instead of polymers of acrylic acid or copolymers of acrylamide and acrylic acid, many other pH sensitive controlled release systems known in the art (Robinson and Lee, 1987) can be used. Examples include soluble anionic cellulose systems, or insoluble crosslinked anionic cellulose systems; Or soluble anionic polymers or insoluble crosslinked anionic polymers derived from any common acrylic acid polymers and / or derivatives thereof. Such crosslinked insoluble polymers are preferred in that they swell and are not easy to metabolize.

The controlled release system preferably includes a pH sensitive crosslinked absorbent pill that is gelled upon wetting.

The present invention also provides an animal feed composition containing the antimicrobial agent and feed of the present invention. The antimicrobial agent is preferably provided here in an amount in the range of 0.0001 to 25% of the total feed composition, particularly preferably in an amount in the range of 0.0001 to 5% of the total composition.

In another preferred embodiment, the antimicrobial agent of the present invention may be formulated in a form for use in addition to drinking water of an animal.

Such an antimicrobial agent of the present invention is preferably administered in an amount in the range of 0.05 to 5000 mg / kg / day, more preferably in the range of 0.05 to 50 mg / kg / day.

Examples of inert carriers suitable for use in the compositions for administering the antimicrobial agents of the invention include water, olive oil, peanut oil, sesame oil, sunflower oil, safflower oil, arachis oil, coconut oil, liquid paraffin, ethylene glycol, propylene glycol, Polyethylene glycol, ethanol, propanol, isopropanol, glycerol, fatty alcohols, triglycerides, polyvinyl alcohols, partially hydrolyzed polyvinyl acetates and mixtures thereof.

Solid forms for oral or rectal administration may include pharmaceutically or veterinary acceptable binders, sweeteners, disintegrants, diluents, flavors, coatings, preservatives, lubricants and / or time delay agents. Suitable binders are acacia gum, gelatin, corn starch, tragacanth gum, sodium alginate, carboxymethylcellulose or polyethylene glycol. Suitable sweeteners are sucrose, lactose, glucose or flavonoid glycosides such as neohesperidine dihydrochalcone. Suitable disintegrants include corn starch, methylcellulose, polyvinylpyrrolidone, xanthan gum, bentonite, alginic acid or agar. Suitable diluents are lactose, sorbitol, mannitol, dextrose, kaolin, cellulose, calcium carbonate, calcium silicate or dicalcium phosphate. Suitable flavoring agents include peppermint oil, presser oil, cherry, orange or raspberry flavors. Suitable coatings include polymers or copolymers of acrylic acid and / or methacrylic acid and / or esters thereof, and / or amides, waxes, fatty alcohols, zein, shellac or gluten thereof. Suitable preservatives are sodium benzoate, vitamin E, α-tocopherol, ascorbic acid, methylparaben, propyl paraben or sodium hydrogen sulfite. Suitable lubricants are magnesium stearate, stearic acid, sodium oleate, sodium chloride or talc. Suitable time delay agents are glyceryl monostearate or glyceryl distearate.

Suspensions for oral or rectal administration may further comprise dispersants and / or suspending agents. Suitable suspending agents are sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, sodium alginate or cetyl alcohol. Suitable dispersants include lecithin, polyoxyethylene esters or fatty acids such as stearic acid, polyoxyethylene sorbitol mono- or di-oleate, -stearate, or -laurate, polyoxyethylene sorbitan mono- or di-oleate , -Stearate or -laurate.

The antimicrobial composition may further comprise one or more emulsifiers. Suitable emulsifiers include dispersants as described above or natural gums such as acacia gum or tragacanth gum.

Compositions for administration in the methods of the invention include methods known in the art (eg, veterinary and pharmaceutical composition arts) of preparing the compositions, such as blending, grinding, homogenizing, suspending, dissolving, emulsifying, dispersing and Where appropriate, the polymer may be prepared by mixing with selected excipients, diluents, carriers and auxiliaries.

For oral administration, pharmaceutical or veterinary compositions may be used in tablets, lozenges, pills, troches, capsules, elixirs, powders (including freeze-dried powders), solutions, granules, suspensions, emulsions, syrups and tinctures. It may be in the form. Sustained release or delayed release may be prepared, for example, in the form of coated particles, multilayer tablets or microgranules.

In general, poly (2-propeneal, 2-propene acid) is preferably prepared from poly (2-propeneal) by oxidation of its solid in air. Specifically, the poly (2-propeneal) polymer may first be heated to a temperature between 80 and 110 ° C., mainly under anhydrous state. More preferably, the polymer is first heated to about 85 ° C. The poly (2-propeneal, 2-propene acid) is preferably heated in alcohol for a period in the range of 1 hour to 1400 hours, more preferably in a range of 1 hour to 60 hours.

According to the invention, there is further provided a preservative compound or composition comprising the antimicrobial agent of the invention.

According to the present invention there is also provided a fungicide or preservative compound or composition comprising the antimicrobial agent of the invention.

According to a further aspect of the present invention, the present invention provides a composition for treating gastrointestinal diseases, including the antimicrobial polymer as described above and additionally a chemotherapeutic agent, wherein an additional chemotherapeutic agent is adsorbed on the antimicrobial agent.

The adsorption can generally reduce the membrane penetration of additional chemotherapeutic agents. Chemotherapeutic agents suitable for use in the present examples show a significant reduction in membrane penetration when mixed with polymeric antimicrobials. Preferably penetration is inhibited by at least 50% proportion.

Chemotherapeutic agents useful for use in this aspect of the invention include antibiotics for treating gastrointestinal diseases and anticancer agents for treating gastrointestinal cancer.

Chemotherapeutic agents used in conjunction with polymeric antimicrobials reduce membrane penetration of chemotherapeutic agents to reduce systemic side effects and provide more targeted treatment. In most cases the odor is also reduced.

Examples of chemotherapeutic agents for treating gastrointestinal diseases include antibiotics and anticancer agents.

Examples of antibiotics that can be used with the antimicrobial polymers are tetracycline, penicillin, aminoglycosides, sulfa drugs, cephalosporins and nitrofuran.

This antibiotic may be a conventional antibiotic used to treat infections of the gastrointestinal tract.

Examples of anticancer agents that can be used with the polymer antimicrobial agents of the present invention include alkylating agents, metabolites, anticancer antibiotics, plant alkaloids, hormones and other anticancer agents, specifically anticancer agents containing only carbon, hydrogen and oxygen.

The compositions of the present invention comprise phenol (preferably 0.1 to 10% by weight), isothiazolinone (preferably 0.001 to 1%), alkyl parabens (preferably 0.02 to 2%) and lower alcohols (preferably 20 To 99%), may further comprise one or more antimicrobial agents such as selected from the group consisting of: Here, the content is in weight units based on the weight of the composition.

It was found that the derivatives of poly (2-propeneal, 2-propene acid) used in the method of the present invention have significantly increased stability compared to the poly (2-propeneal, 2-propene acid) polymer. As evidenced by the loss of antimicrobial activity of the poly (2-propeneal, 2-propene acid) compositions, some of the instability of these compounds has been documented in the prior art, so the inventors have found that "accelerated curing at elevated temperatures, i. ". Surprisingly, an elevated temperature of 40 ° C. “curing” poly (2-propeneal, 2-propenoic acid) in aqueous or polyethylene glycol aqueous solution not only delays the decrease in antimicrobial activity, but in fact poly (2-propeneal, 2- Propenoic acid) was increased (see Example 2 (a) and (b)). This finding is a totally contradictory result, unexpected from the perspective of the prior art, which predicts that a rise in temperature will lead to "accelerated curing", ie, an accelerated loss of antimicrobial activity.

In this specification, a method of forming a new form of the polymer comprising poly (2-propeneal, 2-propene acid) to provide increased antimicrobial activity is referred to as "overactivation" and the resulting polymer is referred to as "overactivation polymer." "It was called.

Even more surprisingly in view of the prior art, the inventors have found that overactivation in aqueous polyethylene glycol solution is promoted by basic conditions followed by acidic conditions. In addition, excessive activation was also promoted by heat and moisture.

Overactivation is facilitated by the presence of polyethylene glycol or polyols or alkanols, since the presence of polyethylene glycol or polyols or alkanols protects and stabilizes the carbonyl group of the polymer from alkali decomposition by reaction with canniza through acetal formation It seems to be because it is made.

Another advantage of overactivation is that overactivation reduces or eliminates acrolein contaminants that are the source of tissue and skin irritation.

Importantly, overactivation is readily available in the form of any aqueous test solution as a result of increased hydrophilicity of the polymer, as evidenced by the expired Australian patent application AU-A-11686 / 95 ("11686/95"). It is an additional reaction that is quite different from any increase in antimicrobial activity that can only be caused by more than one polymer. The inventors have found that substantial additional antimicrobial activity occurs illustratively as a result of repeating the same method described in 11686/95 and then overactivating the partially soluble polymer. It should be noted that even the overactivation did not completely dissolve the polymer of 11686/95, in contrast to the overactivation which first begins by heating the polymer between 80 and 85 ° C.

The optimal time to overactivate the poly (2-propeneal, 2-propene acid) solution is inversely proportional to temperature. In particular, when used in the presence of a hydroxyl solvent and / or base, and then acidified, the transient activity may be performed by curing at room temperature, but this may require a too long time to be impractical.

We have found active ingredients in overactive polymers as described herein suitable for gastrointestinal treatment, preservatives in aqueous products or methods and fungicides or preservatives with the benefit of enhanced antimicrobial activity. In addition, the inventors have discovered that the antimicrobial activity of such fungicides or preservatives increases when their pH increases, such as above pH 6.

A common feature of the present invention is that the group capable of hydrophobic interaction is attached to the antimicrobial agent of the present invention via hemiacetal / acetal formation or by adsorption to increase the antimicrobial activity.

Hereinafter, the present invention will be described in detail with reference to various embodiments, but the scope of the present invention is not limited thereto.

Sterilization test

The sample was dissolved with 1% by weight aqueous sodium bicarbonate to achieve the required concentration (unless otherwise indicated, the polymer is 0.125% by weight). After weighing 19.9 g of the diluted sample into a sterile bottle, 0.1 ml of 10 ⁷ to 10 ⁸ cfu of Ps. Aeruginosa (Ps. Aeruginosa) was inoculated and mixed. At specific time intervals 1 ml of inoculation sample was added to 9 ml of Letheen broth and vortexed. Ten-fold serial dilutions were plated and plated. Next, trypticase soy agar was injected. Incubated at 37 ° C. for 3 days.

Example 1

This example illustrates a process for producing poly (2-propeneal, 2-propene acid) by oxidizing a solid acrolein polymer in air. This poly (2-propeneal, 2-propene acid) production method is a preferred method for producing starting materials for use in the process of the present invention. Water (720 ml, at room temperature of about 20 ° C.) and acrolein (60 g; freshly distilled, with 0.25 w / w% hydroquinone added) are added to an open beaker in a fumigation cupboard, very strongly mechanically Stirred. Then, pH was adjusted to 10.5-11.0 by adding 0.2M aqueous sodium hydroxide solution (21.4 ml).

This solution immediately turned to the typical yellow color of the hydroquinone anion, disappeared within minutes, and the clear solution became milky white.

After about 1 minute, downy white polymer began to precipitate and a complete precipitate appeared within 15 to 30 minutes. The precipitate was filtered and washed with water (250 ml), then dried at room temperature for 2 days (yield 25 g), then spread in a thin layer in a glass petri dish and heated at 40 ° C./8 h. The heating was then continued in the following order: 50 ° C./15 hours; 65 ° C./4 hour; 75 ° C./18 hours; 84 ° C./24 hours.

It is contemplated that the method can be increased in a sealed container by adding acrolein stepwise and then dried faster (compare Example 10).

In general, a solution of the final material, poly (2-propaneal, 2-propene acid), is added to 2 g of the polymer in 1 w / w% aqueous sodium carbonate solution (100 ml), stirred for 15 to 30 minutes, and then diluted as necessary. It was prepared by. This solution was very clear in contrast to the dissolution attempted alternatively using a polymer from Example 5 of 11686/95.

Example 2

This example illustrates acetal formation from poly (2-propeneal, 2-propene acid).

(a) 5 g of poly (2-propaneal, 2-propene acid) were dissolved in 64 g of polyethylene glycol ("PEG") 200 and mixed with 31 g of 0.71 w / w% sodium carbonate solution. A portion of this solution (apparent pH = 5.8) is kept at room temperature

(b) The rest of sample (a) was heated to 60 ° C. for 12 or 25 days.

The samples of (a) and (b) were diluted with 1w / w% sodium bicarbonate and subjected to a sterilization test with a polymer of 0.125w / w% concentration. Surprisingly, it was confirmed that the samples subjected to the "accelerated cure" exhibited improved antimicrobial activity, the results of which are shown in Table 1 for reference.

Cfu / ml ^* (Pseudomonas aeruginosa) sample 0 min 10 minutes 15 minutes 30 minutes 60 minutes 25 days at room temperature 7.8 x 10 ⁶ 4.1 x 10 ⁶ 6.1 x 10 ⁵ 9.8 x 10 ⁴ <10 12 days at 60 ℃ 7.7 x 10 ⁶ 1.4 x 10 ⁶ 9.8 x 10 ³ <10 <10 25 days at 60 ℃ 1.0 x 10 ⁷ 1.3 x 10 ⁶ 6.6 x 10 ⁴ <10 <10 ^* Colony forming unit / ml

1 g of poly (2-propeneal, 2-propenoic acid) was dissolved in 200 ml of 0.1w / w% Na ₂ CO ₃ and left overnight. To this was added sodium lauryl sulfate at a concentration of 0.05 w / w% and the solution was acidified to pH 5.9 with HCl. A portion of this solution was stored at room temperature and 60 ° C. respectively. A sterilization test was conducted with 0.125 w / w% polymer solution prepared using ₁ w / w% NaHCO ₃ as diluent. As can be seen from Table 2 for reference, the "cured" sample showed a remarkable performance improvement:

Cfu / ml ^* (Pseudomonas aeruginosa) sample 0 min 10 minutes 15 minutes 30 minutes 20 days at room temperature (RT) 9.0 x 10 ⁶ 5.1 x 10 ⁵ 6.8 x 10 ² <10 7 days at 60 ° C + 13 days at RT 9.0 x 10 ⁶ 1.2 x 10 ² <10 <10 ^* Colony forming unit / ml

(c) A 5 w / w% solution of the overactivated polymer was prepared as in Example (2a) except that PEG 1000 was used instead of PEG 200. A portion of this solution was treated with concentrated NaOH to pH 8.1. This sample was heated to 60 ° C. and used for sterilization test. As such, the samples exposed to more basic conditions unexpectedly showed excellent sterilization performance as can be seen with reference to Table 3 below:

Cfu / ml ^* (Pseudomonas aeruginosa) sample 0 min 5 minutes 10 minutes 15 minutes 30 minutes 12 days at pH 5.8, 60 ° C 3.8 x 10 ⁶ 2.7 x 10 ⁶ 1.5 x 10 ⁶ 3.3 x 10 ³ <10 7 days at pH 8.1, 60 ℃ 90 x 10 ⁶ - 10 <10 <10 17 days at pH 8.1, 60 ° C 8.3 x 10 ⁶ 3.3 x 10 ⁵ 1.3 x 10 ² <10 <10 ^* Colony forming unit / ml

Example 3

This example examines the product obtained by reacting poly (2-propeneal, 2-propene acid) with polyethylene glycol.

Acetal present in the polymer of Example 2 (b) was measured by irradiating the proton ( ¹ H) and carbon ( ¹³ C) NMR spectrometer with the remaining solid residue after dialysis and concentration of the polymer solution. Dialysis removes all materials with molecular weight less than 1000. Table 4 presents proton ( ¹ H) and carbon ( ¹³ C) NMR data. As can be seen in Table 4, the nuclear magnetic resonance spectroscopy of the residue showed peaks at δ3.58 and 3.56 in the ¹ H nuclear magnetic resonance spectrum and peaks at δ71.62, 69.48 and 60.25 in the ¹³ C nuclear magnetic resonance spectrum. . These peaks indicate that the polyethylene glycol unit was attached as acetal.

domain Carbonyl Alkenyl Vinyl methine Vinyl methylene Acetal Oxyalkyl PEG Alkyl ¹ Hδ 9.5-8.5 7.3-6.0 6.0-5.5 5.5-5.0 5.0-4.0 4.0-3.0 3.58, 3.56 3.0-0.5 ¹³ Cδ 185-175 140-105 105-95 80-55 71.62, 69.48, 60.25 55-15

600 MHz ¹ H- and 125 MHz ¹³ C-nuclear magnetic resonance spectral data in 1w / w% Na ₂ CO ₃ and D ₂ O analyzing solid residues of the overactivated polymer after dialysis and concentration

Example 4

(a) A 5w / w% solution of polymer was prepared similarly to Example 2 (a) except changing the proportion of PEG 200 with an apparent pH of 5.7 and a range of overactivities.

The sample was heated to 60 ° C. and the stability over time was monitored. It was considered that physical stability was not achieved when precipitation or gelation occurred. UV measurements were performed on 0.01 w / w% polymers prepared with 1 w / w% sodium carbonate solution. The decrease in absorption at 268 nm: 230 nm was considered to be equivalent to the decrease in chemical stability. The results are shown in Table 5:

Furtherance A B C D PEG200 (% by weight) 0 50 64 95 Physical stability time A B C D 4 days, 60 ℃ failure Pass Pass Pass 11 days, 60 ℃ failure failure Pass Pass Chemical stability Ratio = 260-270 peak absorbance / 228-235 peak absorbance time A B C D 0 days, 60 ℃ 1.38 1.41 1.43 1.46 4 days, 60 ℃ 0.98 1.04 1.21 1.27 11 days, 60 ℃ - 0.97 1.03 1.09 18 days, 60 ℃ - 0.89 0.92 1.04 25 days, 60 ℃ - - 0.84 1.04

Both physical and UV spectral results demonstrated that PEG had a positive effect on stability, and higher PEG content resulted in greater physical and chemical stability.

(b) The following solutions A and B were dissolved in 196 g of 1w / w% sodium bicarbonate in 4 g of poly (2-propaneal, 2-propene acid), and then adjusted to pH 7 (A) and 5.5 (B) with dilute hydrochloric acid. So Prepared. Solution C was prepared by dissolving 50 g of poly (2-propenal, 2-propenic acid) in PEG 200 (640 g) at 65 ° C to 70 ° C. Then, a solution obtained by dissolving 4 g of sodium carbonate in water (306 g) was added and found to have an apparent pH of 7 and to 5.5 after 31 days of treatment.

All samples were stored at 40 ° C. Sterilization tests were performed with samples containing 0.125 w / w% of polymer equivalents at various time intervals. The results are shown in Table 6:

Time in days at 40 ℃ Time to complete kill of <10 cfu / ml (minutes) (Pseudomonas aeruginosa) Solution A Solution B Solution C 0 30 30 30 7 30 60 - 14 - - 10 31 60 60 10

Example 5

1 g of poly (2-propeneal, 2-propenoic acid) was heated to 60 ° C. for 3 days in anhydrous or wet sealing chamber. The anhydrous polymer solution and the humidified polymer solution were each prepared at 0.125 w / w% (corrected for moisture content) and evaluated through sterilization tests.

Cfu / ml ^* (Pseudomonas aeruginosa) 0 min 5 minutes 10 minutes 15 minutes 30 minutes 60 minutes Polymer (anhydrous) 4.9 x 10 ⁶ - 7.6 x 10 ⁵ 5.9 x 10 ⁴ 1.2 x 10 ² <10 Polymer (humidification) 1.1 x 10 ^{7 6} x 10 ⁶ 3.4 x 10 ³ 3.7 x 10 ³ <10 - ^* Colony forming unit / ml

These polymers exhibit carbonyl and / or carboxyl absorption at IRs between 1700 and 1730 cm ⁻¹ , exhibit carbonyl groups (eg, when using Schiff reagent), Mw is about 10000 and Mn is about 5000: titration results carboxyl groups Was found to be about 5 mol%. These variables are similar (but not identical) to those of poly (2-propenal, 2-propenoic acid).

Example 6

In two replicate experiments, polymer samples were prepared and then dissolved in ethane-diol as described in Example 5 of 11686/95. Half of this material was further heated at 80 ° C. for 24 hours (therefore the solubility of the aqueous medium remained incomplete). Standard antibacterial tests were used to compare the antimicrobial activity of the samples. The heat treated, ie, superactivated, samples all showed a marked improvement in antimicrobial activity as shown in Table 8:

Cfu / ml ^* (Pseudomonas aeruginosa) Solution treatment First number 5 minutes 10 minutes 15 minutes 30 minutes (1) no treatment 4.6 x 10 ⁶ 5.7 x 10 ⁵ 2.9 x 10 ² <10 <10 (2) no treatment 4.6 x 10 ⁶ 4.2 x 10 ⁵ 1.5 x 10 ² 10 <10 (1) 80 ° C, 24 hours 4.6 x 10 ⁶ 3.7 x 10 ⁶ <10 <10 <10 (2) 80 ° C, 24 hours 4.6 x 10 ⁶ 8.0x10 ⁵ <10 <10 <10 ^* Colony forming unit / ml

Example 7

50 g of poly (2-propeneal, 2-propene acid) were dissolved in PEG200 (640 g) between 65 and 70 ° C. Then, an aqueous solution in which sodium carbonate (4 g) was dissolved in water (306 g) was added. The samples were divided and left at room temperature or heated at 80 ° C. for 24 hours. Over time, the acrolein content of the solution was determined by reversed phase HPLC and the results are shown in Table 9:

20 ℃ storage days Acrolein content (ppm) Overactive Not overactivated 0 274 144 7 - 126 16 34 103 30 13 80

Example 8

Poly (2-propeneal, 2-propene acid) solutions were prepared as in Example 7, and treated at temperatures of 40 ° C., 60 ° C., 80 ° C., 100 ° C. and 115 ° C. at various times. Samples were treated with standard sterilization tests to confirm the increase in mortality and the results are shown in Table 10:

Transient activation temperature (℃) Optimum time range (hours) Total kill time (minutes) Room temperature > 1400 <10 40 1400 <10 60 120-170 <10 80 16-24 <10 100 4-7 <10 115 1-3 <10

The time required for overactivation was observed to be inversely proportional to temperature. All polymer solutions obtained from the overactivation process were fully miscible in the aqueous solvent at all ratios.

Example 9

(a) 540 g of poly (2-propaneal, 2-propene acid) was dissolved in 2304 g of PEG200 at 65 ° C. and then mixed with a solution of 43.2 g of sodium carbonate dissolved in 712 g of water. The solution was then heated at 100 ° C. for 4 hours and 36 g of sodium lauryl sulfate, 7 g of ECOTERIC T20 (nonionic surfactant) and 2 g of lemon flavor were added. This preparation (pH6) is diluted 1:30 with hard water and Staphylococcus aureus (particularly Gram-positive bacteria that are significant for infections in hospitals) and Salonella choleraesus (particularly infections in food manufacturing). Gram-negative bacteria, which are significant in relation to the above, were administered according to the Association of Agricultural Chemists Official Methods of Analysis (1995) 991.47, 991.48, and Hard Surface Carrier Test Method. The results are shown in Table 11:

microbe Positive test tube result S. aureus 2/60 Pass S. Choleraesus 1/60 Pass

Increasing the pH of this formulation resulted in increased antibacterial activity as confirmed by the sterilization test. The results are presented in Tables 12 (a) and 12 (b):

pH 10 minutes (cfu / ml) 20 minutes (cfu / ml) 30 minutes (cfu / ml) 45 minutes (cfu / ml) 60 minutes (cfu / ml) 5.6 2.8 x 10 ⁵ 4.4 x 10 ⁴ 2.3 x 10 ³ 20 <10 7.2 2.7 x 10 ³ <10 <10 <10 <10 8.9 3.2 x 10 ³ <10 <10 <10 <10 10.5 1.1 x 10 ² <10 <10 <10 <10

Active against Staphylococcus aureus

Initial bacterial count, 3 × 10 ⁶ cfu / ml; Polymer 350ppm

pH 10 minutes (cfu / ml) 20 minutes (cfu / ml) 30 minutes (cfu / ml) 45 minutes (cfu / ml) 60 minutes (cfu / ml) 5.6 2.9 x 10 ⁵ 8.6x10 ⁴ 6.2 x 10 ² 40 <10 7.2 5.8 x 10 ⁵ 9.1 x 10 ⁴ 4.3 x 10 ³ <10 <10 8.9 9.5 x 10 ⁵ 8.2 x 10 ⁴ 4.6 x 10 ² <10 <10 10.5 1.1 x 10 ² 3.0 x 10 ³ <10 <10 <10

Activity against Pseudomonas aeruginosa

Initial bacterial count, 3.7 × 10 ⁶ cfu / ml; Polymer 350ppm

(b) 1200 g of poly (2-propaneal, 2-propenoic acid) was dissolved in 7680 g of PEG200 at 60 ° C., and then a solution of dissolving 96 g of Na ₂ CO _{3 in} 3024 g of water was added. This solution was heated at 100 ° C. for 6 hours.

This formulation was added three times a week at a concentration of 300 ppm (30 ppm of polymer) to the collector of the induction draft cooling tower. Feeding was carried out in the evening so that the contact time before the recommended operation was 8 to 12 hours. As a result, the residual concentration was estimated to be 1/2 every 3-6 hours of operation. Circulating water had an average temperature of 27 ° C., a pH of 8.5, and a conductivity of 3000 µS. The number of microorganisms was measured and compared to the same tower in the vicinity where daily biodispersant was supplied. The results are shown in Table 13.

Cfu / ml ^* Processing time (days) Treated tower Control tower One 2.4 x 10 ³ 1.1 x 10 ⁷ 2 2.0 x 10 ³ 1 x 10 ⁶ 3 3.3 x 10 ³ - 4 2.5 x 10 ³ - 14 6.1 x 10 ⁴ 2.6 x 10 ⁶ 15 5.1 x 10 ⁴ 1.1 x 10 ⁶ 16 5.1 x 10 ⁴ 4.9 x 10 ^{6 *} Colony forming unit / ml

This data indicates that the treatment program maintains the number of microorganisms within the criteria of AS / NZ Standard 3663.3 (Int): 1998 and contains a biodispersant (which is observed to be particularly unsuitable for the test conditions required in very hot summer months). It has been shown to maintain a lower microbial count than the number of microorganisms present.

Example 10

10 (a) Comparative Example

This example illustrates the preparation of an acrolein polymer in which the method of the present invention is not used.

1.0 0.8w / w% Sodium Hydroxide

9.90 kg of deionized water was added to a 10 L stainless steel barrel, and 0.08 kg of sodium hydroxide was added to the water, followed by stirring until dissolved.

2.0 polymerization

100.1 kg of deionized water was placed in a 200 L stainless steel keg, and 4.99 kg of 0.8 w / w% sodium hydroxide solution was added to the keg. The solution was equilibrated to 15-20 ° C. At the same time, 20 kg of acrolein monomer was added and the remaining 0.8w / w% sodium hydroxide solution was added to this 200L barrel at a constant rate over 1 hour so that the pH was maintained at 10.5 to 11.0 and the temperature did not rise above 30 ° C. The polymerization was continued for 90 minutes.

3.0 wash

The polymerization mixture was filtered / centrifuged and the polymer was washed with deionized water until the pH of the wash water was below 7.0. The approximate yield was 8 kg.

4.0 drying

The polymer was dried in air and then heated in an oven in the following order.

step time Temperature One 2 hours 25 ℃ 2 1 hours 40 ℃ 3 1 hours 70 ℃ 4 1 hours 75 ℃ 5 2 hours 85 ℃

5.0 melting

400 L of water was added to a 500 L barrel, and then 4 kg of sodium carbonate was added and stirred until dissolved. 8 kg of heated anhydrous polymer was added slowly and stirred for 30 minutes.

The resulting polymer was found to have an approximate solubility of 90-95% in 1w / w% sodium carbonate.

Example 10 (b)

This example illustrates the preparation of an acrolein polymer in which the polymer of the comparative example is overactivated by the method of the present invention.

1.0 Base Preparation

0.4 kg of sodium carbonate was dissolved in 30.6 kg of water in a suitable vessel and 64 kg of polyethylene glycol 200 was added to this mixing vessel. Agitation was started with a mechanical stirrer and the PEG200 was heated to 65 ± 3 ° C. 5 kg of the anhydrous acrolein polymer of Example 10a was added to this PEG200 and stirred until a homogeneous mixture was obtained.

Note: Solids may not be completely dissolved at this stage.

Thus, sodium carbonate solution is slowly added to the glycol mixture at a rate such that the pH of the solution can be maintained in the range from 3.5 to 9.0.

This solution was stirred at 65 ± 3 ° C. for 45 minutes.

Note: The pH should range from 7 to 9 and the temperature should range from 65 ± 3 ° C.

2.0 transient activation

The mixing vessel was covered and heated to 100 ° C. for 4 hours. The resulting polymer was observed to have an approximate solubility in water of 99.5 to 100%.

Example 11

This example examines the antimicrobial activity of the normally activated anhydrous poly (2-propeneal, 2-propene acid) polymer shown in Example 10a and the overactivated acetal derivatives shown in Example 10b.

The chicks were treated with each antimicrobial agent and compared with the control group according to the following procedure:

Twenty newborn chicks (line 53) newborns were purchased for each trial at the commercial hatcheries, weighed, and randomly divided into neighboring females in separate nurseries. Male and female chicks evenly distributed. Water and feed were to be eaten from time to time. The feed was a commercially available crumble (Chick Starter, Milne Feeds: 18% crude protein) containing a coccidium inhibitor (125 ppm dinitolamide).

Ten chicks were administered via a fixed water feeder for 14 days with a formulation in which 0.1w / w% of the normally activated antimicrobial agent of Example 10a was added to water. Dosage rate was 30 mg / kg / day. The other ten chicks served as controls.

Two groups of chicks were weighed on days 0, 4, 7, 11 and 14. At the end of this test all chicks were euthanized and the treated chicks were post-dissected. Thoracic and abdominal cavity were thoroughly and thoroughly investigated.

result

Work Control (average body weight g) Treatment group (average body weight g) Difference(%) 0 42.5 42.5 0 4 65.0 72.5 11.5 7 98.5 98.0 -0.5 11 145.5 148.5 2.4 14 185.5 200.5 8.1

Weight gain during normal activated antimicrobial treatment of Example 10a

The treatment group showed some weight gain on the last 14 days compared to the control.

At the end of the trial, no overall clinical or pathological signs of toxicity were observed in the overall investigation of the chicks treated at the dissection.

Work Control (average body weight g) Treatment group (average body weight g) Difference(%) 0 42.5 42.5 0 4 62.5 67.5 8 7 97.5 103 6 11 130 145 11.5 14 178 219 23

Weight gain during the overactivated acetal antimicrobial test of Example 10b

At the end of the trial, the rest of the chicks showed no signs of clinical or pathological toxicity in the aggregate investigations of both groups.

conclusion:

Compared to the control group (χ ² ; P <0.015), there was a significant difference in weight gain in the treatment group. At the end of the test, the treatment group was 23% heavier than the control group.

Significant improvement in weight gain of the overactivated acetal derivatives compared to normal activated poly (2-propeneal, 2-propenoic acid) in the control and the following examples demonstrated a significant improvement in the intestinal antibacterial activity of the acetal derivatives. It is.

Example 12

This example evaluates the polymeric antimicrobial agent of Example 10b to inhibit weaning pig E. coli (PWC) under field conditions.

Way:

146 young pigs (organic pigs) that have been orally vaccinated, auto-vaccinated or untreated with over-activated antimicrobial agents of various compositions via feed, water or APRALAN® (Elanco) Large pig farms were treated with weaning-related stress.

The response of diarrhea, weight gain and lethality development was assessed and reported in Table 15.

Number of overactivated polymer antibacterial groups ¹ Death rate [% of deaths] ² Diarrhea Days [Diarrhea Days: 1-2] ³ Average bowel movements ⁴ Overactivated Polymer Antibacterials One 3.33 1.79 [F; P <0.0001] 0.14 [F; P <0.0001] 2 0 1.17 [F; P <0.0001] 0.11 [F; P <0.0001] Untreated control 3 16.67 [F; P <0.05] 3.77 [F; P <0.0001] 0.41 [F; P <0.0001] Apralan® Group 4 13.33 [F; P <0.05] 3.89 [F; P <0.0001] 0.38 [F; P <0.0001] Vaccination Group 5 6.67 3.10 [F; P <0.0001] 0.30 [F; P <0.0001]

week:

1. Processing Coding:

i. Group 1 = 0.1w / w% overactivated polymer antimicrobial in feed

ii. Group 2 = 0.02w / v% overactivated polymer antimicrobial in water

2. Percentage of groups killed by PWC during the trial

3. Average number of days in each pig in the group with one or two bowel movements; Defecation is a measure of diarrhea.

4. The total number of bowel movements divided by the number of samples observed per pig.

5. F = Fisher's accuracy test.

conclusion:

This field test positively reflects the efficacy of acetal derivatives of poly (2-propenal, 2-propenic acid) for use in young piglets in commercial pig farms after weaning β-hemolytic E. coli The emphasis is on the same point:

1. Mortality rate:

Lower mortality seen in overactive polymer antimicrobial treated groups in the untreated, vaccinated or Apralan® group

2. Diarrhea days:

Polymeric antimicrobial treatment group overactive in untreated group, vaccinated group or Apralan® group [F; Significantly lower diarrhea days at P <0.0001]

3. Defecation frequency:

More overactivated antimicrobial treatment group [F; Significantly lower mean defecation frequency at P <0.0001]

Example 13

This example investigates the effect of using a specific additive with the antimicrobial agent of the present invention.

Way:

Dosing cultures were incubated overnight before the total viable count (TVC) of these cultures was measured.

This sample was serially diluted 1: 1 with sterile general saline (5 ml). If the test sample contained EDTA, sterile hard water (SHW) was used as the diluent.

One part of each culture incubated overnight was diluted with 9 parts of diluent. This diluted solution was used as inoculum.

100 μl of this diluted overnight culture was inoculated into each sample dilution (1 culture per test tube). Mix well.

Each test tube was incubated at 37 ° C. for ≦ 24 hours (A. Niger was incubated at 28 ° C. for ≦ 24 hours).

1 ml was taken from each test tube and inoculated twice in 9 ml of restored medium and mixed well (Restore medium: nutrient liquid medium + 3% Tween 80 (NBT) for polymer antibacterial and EDTA; nutrient liquid medium + 3% for glutaraldehyde) Tween 80 + 0.1% ammonia (NBTA), rettenx (LB) for methyl parabens).

Samples were incubated at 37 ° C. for additional ≦ 48 hours (A. Niger for 28 days at 28 ° C.).

Each tube was examined for proliferation. All test tubes were secondary cultured in selection agar medium and incubated at 37 ° C. for ≦ 24 hours (A. Niger for 28 days at 28 ° C.). Proliferation in selection agar medium was considered to be the proliferation of test organisms.

As a positive control, it was incubated using a 5 ml dilution inoculated into the culture, recovered and confirmed.

As a negative control, uninoculated 5 ml dilutions were used to incubate, and then recovered.

result:

The results are shown in Table 16 below.

MKC and Synergy Rates of Selected Preservatives

Note: All ratio values are in the ratio of polymer antimicrobial to preservative.

Culture: Culture

Inoculum: Inoculum

A.niger: A. Niger

C. albicans: C. albicans

E.coli: Escherichia coli

P.aeruginosa: P. aeruginosa

S.aureus: S. aureus

Code Description:

1) Polymer antibacterial 0.025w / w%

2) polymer antibacterial 0.2w / w%

3) Glutaraldehyde 0.025w / w%

4) Polymer antibacterial 0.025w / w% + glutaraldehyde 0.025w / w%

5) Polymer antibacterial 0.2w / w% + glutaraldehyde 0.025w / w%

6) polymer antibacterial 0.2w / w%

7) EDTA 0.1w / w%

8) Polymer antibacterial 0.2w / w% + EDTA 0.1w / w%

9) 0.2w / w% polymer antibacterial (in 80w / w% glycerol)

10) Methylparaben 1 w / w% (in 80 w / w% glycerol)

11) 0.2w / w% polymer antibacterial (in 80w / w% glycerol) + 1w / w% methylparaben (in 80w / w% glycerol).

Culture Glutaraldehyde EDTA Methyl paraben A. Niger <0.6 <0.5 0.2 C. albicans 0.3 0.1 0.5 E. Coli 0.5 <0.3 0.3 P. aeruginosa 0.6 0.4 0.2 S. aureus 0.3 <0.7 0.2

Synergy Rate by Polymer Antibacterial Agents

Note: Synergy <1.0, Additive = 1.0, Antagonism> 1.0

SI = CD / A + CE / B

A = MKC Polymer Antibacterial

B = MKC Preservative

C = MKC Mixture

D = A: B ratio

E = B: A ratio

conclusion:

Acetal derivatives of poly (2-propenal, 2-propenoic acid) include glutaraldehyde, EDTA and methyl for A. Niger, C. albicans, E. coli, P. aeruginosa, S. aureus. The action of each paraben was elevated.

Example 14

This example demonstrates the activity of the antimicrobial agent according to the invention together with the antimicrobial agent under the trade name "Dettol".

Samples were serially diluted in a ratio of 1: 1 using normal sterile saline (5 ml).

Each sample dilution was inoculated with 100 μl of diluted overnight culture (1 culture per test tube). Samples were mixed well.

These samples were incubated at 37 ± 2 ° C. for ≦ 24 hours (in vitro inoculated with Aspergillus niger was incubated at 28 ± 2 ° C. for ≦ 24 hours).

1 ml was taken from each test tube and secondaryly cultured in 9 ml of reconstituted medium and mixed well (Sabour name + 3% Tween 80 (SABT) for NBT, A. Niger).

This was incubated at 37 ± 2 ° C. for ≦ 48 hours (A. Niger was incubated at 28 ± 2 ° C. for 5 days).

The turbidity (growth) of the restored medium was measured, and the growth was confirmed by plating on the selected agar medium.

Culture Inoculation Approx. (Cfu / ml) Polymer Antimicrobial MKC Range (ppm) One 2 3 4 5 A. Niger 4.1 x 10 ⁷ 2000-500 N / A 1000 300 N / A 125: 150 C. albicans 5.2 x 10 ⁶ 100-250 N / A 500 75 N / A 62.5: 75 E. Coli 7.6 x ^{10 8} 125-31 125 N / A 75 31:75 N / A P. aeruginosa 1.8 x ^{10 9} 250-62.5 N / A 250 600 N / A 125: 150 S. aureus 2.0 x ^{10 9} 31-7 N / A 15 75 N / A 15:19

MKC results (ppm) of overactivated polymeric antimicrobials and / or "Dettol"

Description

1) 0.1w / w% polymer antibacterial

2) 0.2w / w% polymer antibacterial

3) "Dettol" 4.8w / v% (1:20 dilution)

4) 0.1w / w% Polymer Antibacterial + "Dettol" (1:20 dilution)

5) 0.2w / w% Polymer Antibacterial + "Dettol" (1:20 dilution)

Culture Synergy rate (SI) A. Niger 0.3 C. albicans 0.6 E. Coli 0.6 P. aeruginosa 0.4 S. aureus 0.6

Synergism of Polymer Antibacterials and Dettol

Note: SI> 1-sided antagonist, SI = 1 if additive, SI <1-sided synergist

SI = CD / A + CE / B

A = MKC polymer antimicrobial (ppm)

B = MKC Dettol (ppm)

C = polymer antimicrobial MKC / "Dettol" mixture (ppm)

D = ratio of polymeric antimicrobial to Dettol

E = ratio of "Dettol" to polymeric antimicrobial

Note: The active antimicrobial agent of "Dettol" is Chloroxylenol

conclusion:

The polymer antimicrobial agent of the present invention was found to elevate the action of "Dettol" against E. coli, S. aureus, P. aeruginosa, C. albicans and A. Niger. Table 19 shows that "Dettol" and the polymeric antimicrobial agent work significantly better than when used together as a mixed solution.

Example 15

Preservative of poly (2-propenal, 2-propene acid) [overactivated]

Antimicrobial poly (2-propenal, 2-propene acid) [overactivated] was investigated for preservatives.

The number of bacteria in the subject's hands was measured before and after application of the antimicrobial agent, and then surgical gloves were worn. 4% Chlorhexidine Surgical Scrub, a surgical preservative commonly used to preserve the preservative effect of poly (2-propenal, 2-propenic acid) [overactivated] Orion Retortoris, Perth, Western Australia Product).

A 3w / w% aqueous solution of poly (2-propeneal, 2-propene acid) [overactivated] reduced the reference number of the bacterial population of gloved hands after 3 hours.

A 2w / w% aqueous solution of poly (2-propenal, 2-propenic acid) [overactivated] used with 70% ethanol maintains a reduced bacterial baseline of the gloved hand as 4% chlorhexidine after 3 hours. I was.

3.2w / w% aqueous solution of poly (2-propenal, 2-propenic acid) [overactivated] and subsequent use with 3.1w / w% sodium lauryl sulfate applied to the hand before wearing surgical gloves and subsequent A 4w / w% aqueous solution of poly (2-propenal, 2-propenic acid) [overactivated] in 70% ethanol significantly reduced the bacterial baseline after 3 hours.

These results show that poly (2-propeneal, 2-propene acid) [overactivated] exhibits good residual antimicrobial activity for the continuous inhibition of bacterial counts in sterile conditions during surgery. The addition of 70% ethanol to this formulation helps to rapidly reduce the bacterial count initially.

Example 16

Bactericidal Activity of 0.125w / w% and 0.05w / w% Overactivated Polymers on Control Strains H. Pylori NCTC 11637 at pH 7 and pH 4

First, the in vitro efficacy of the overactivated polymer against H. pylori control strain, H. pylori NCTC11637 was measured. As a variable, two concentrations were selected: one is a 40-fold dilution of the 5% solution of the overactivated polymer prepared in Example 2, which is a hyperactivated polymer at a concentration of 0.125 w / w% similar to the dilution rate above; The other is an overactivated polymer at a concentration of 0.05 w / w%, which is a 100-fold dilution. The pH was chosen as pH 7 and pH 4 similar to the above state.

H. pylori NCTC11637 cultures were grown aerobicly in agar plate for selection at 37 ± 2 ° C. until sufficient growth was observed. Proliferation was aseptically separated from the plates, prepared as a standard turbidity suspension of 10% T as indicated in the Vitek colorimeter and diluted with normal sterile saline. After weighing 19.9 g of the sample, it was inoculated with 100 µl of the culture suspension. 1 ml of the sample was immediately transferred to inactivation / restore medium (Nutrient medium + 3% Tween 80) and then serially diluted. 100 μl of this culture was injected into agar plate for selection and plated using a disposable sterile spreader. The migration step was then repeated at 5, 10, 15 and 20 minute intervals. All plates were incubated microaerobically at 37 ± 2 ° C. until sufficient growth (about 5-7 days). All colonies were counted and subjects declined over time. It was repeated using normal sterile saline as a sample to measure the natural depletion in atmospheric conditions.

Code Description:

Culture 1. Overactive Polymer, pH 7, 0.125w / w%

Culture 2. Overactivated Polymer, pH 7, 0.05 w / w%.

Culture 3. Overactivated Polymer, pH 4, 0.125 w / w%.

Culture 4. Overactivated Polymer, pH 4, 0.05 w / w%.

Culture 5. Normal Sterilized Saline, pH 7.

Culture 6. Normal Sterilized Saline, pH 4.

Culture T = 0 minutes T = 5 minutes T = 10 minutes T = 15 minutes T = 20 minutes One 1.0 x 10 ⁵ 8.6x10 ³ 1.0 x 10 ² <1.0 x 10 ² <1.0 x 10 ² 2 1.6 x 10 ⁵ 2.0 x 10 ³ 4.0 x 10 ² 1.0 x 10 ² <1.0 x 10 ² 3 1.0 x 10 ⁵ 3.2 x 10 ⁴ 1.1 x 10 ⁴ 4.0 x 10 ² <1.0 x 10 ² 4 3.0 x 10 ⁴ 1.7 x 10 ⁴ 1.1 x 10 ⁴ 8.2 x 10 ³ 3.6 x 10 ³ 5 2.0 x 10 ⁵ 1.0 x 10 ⁵ 9.2 x 10 ⁵ 8.0x10 ⁴ 7.9 x 10 ⁴ 6 2.0 x 10 ⁴ 1.4 x 10 ⁴ 8.0x10 ³ 6.0 x 10 ³ 6.0 x 10 ³

Bactericidal Activity of CHEMEQ ^RTM Polymer Antibacterial Agent against H. pylori (NCTC11637)

Note: Number of bacteria per 1 ml of inactivated liquid medium, colony forming unit (cfu)

The results for the control strain (Table 20) showed that the overactivated polymers were effective at pH 7 at 0.125w / w% and 0.05w / w% concentrations, and also at pH 4 and 0.125w / w.

Example 17

In addition to the analysis of Example 16, the other three strains of H. pylori were examined at pH 7 and pH 4: H. pylori 01/303 resistant to clarithromycin and metronidazole; H. pylori SS1, clinical strain isolated from Sydney with high colony of interest in possible animal models; And H. pylori ATCC 700392, a strain obtained from the UK and whose genome has been analyzed.

Culture T = 0 minutes T = 5 minutes T = 10 minutes T = 15 minutes T = 20 minutes Control group H. pylori 11637 2.0 x 10 ⁵ 1.0 x 10 ⁵ 9.2 x 10 ⁵ 8.0x10 ⁴ 7.9 x 10 ⁴ H.Pylori 11637 1.0 x 10 ⁵ 8.6x10 ³ 1.0 x 10 ² <1.0 x 10 ² <1.0 x 10 ² H.Pylori 01/303 3.6 x 10 ⁵ 2.8 x 10 ³ <1.0 x 10 ² <1.0 x 10 ² <1.0 x 10 ² H.Pylori SS1 2.8 x 10 ⁵ 1.4 x 10 ^{3 2.0} x 10 ² <1.0 x 10 ² <1.0 x 10 ² H.Pylori 700392 2.0 x 10 ⁶ 1.3 x 10 ⁶ 1.8 x 10 ⁵ 8.8 x 10 ³ <1.0 x 10 ²

Bactericidal Activity of Overactivated Polymer at 0.125%, pH 7

When treated with 0.125 w / w% overactivated polymer at pH 7, all strains died rapidly, particularly susceptible antibiotic resistant strains to kill within 10 minutes (Table 21). The control strain was untreated.

Example 18

The method of Example 3 was repeated to test the bactericidal activity of 0.125w / w% overactivated polymer (pH 4) for all H. pylori strains.

Culture T = 0 minutes T = 5 minutes T = 10 minutes T = 15 minutes T = 20 minutes Control group H. pylori 11637 2.0 x 10 ⁴ 1.4 x 10 ⁴ 8.0x10 ³ 6.0 x 10 ³ 6.0 x 10 ³ H.Pylori 11637 1.0 x 10 ⁵ 3.2 x 10 ⁴ 1.1 x 10 ⁴ 4.0 x 10 ² <1.0 x 10 ² H.Pylori 01/303 4.2 x 10 ⁶ 1.1 x 10 ⁶ 5.0 x 10 ⁴ 2.0 x 10 ^{4 2.0} x 10 ² H.Pylori SS1 4.9 x 10 ⁶ 1.0 x 10 ⁶ 1.0 x 10 ⁵ 2.8 x 10 ^{4 2.0} x 10 ² H.Pylori 700392 5.5 x 10 ⁶ 1.9 x 10 ⁶ 1.2 x 10 ⁵ 3.6 x 10 ⁴ <1.0 x 10 ²

Bactericidal Activity of CHEMEQ ^RTM Polymer Antibacterials at 0.125% and pH 4

Testing of the overactivated polymer at 4 and 0.125w / w% at similar pH as above resulted in all strains killed within 20 minutes (Table 3). This result is important because the time interval required to kill the H. pylori is less than the transit time (40 minutes to 1 hour). This demonstrates the effect of the overactivated polymer at pH, concentration and time intervals consistent with treating H. pylori infection in the stomach. Control strains were not treated.

Example 19

This example demonstrates the intestinal antimicrobial activity of the acetal derivatives of poly (2-propeneal, 2-propene acid) prepared according to the procedure of Example 10b.

Material and method:

Sixteen weaning pigs (age: 18 days ± 2 days, body weight: 5.5 kg ± 1.0 kg) were purchased from commercial pig farms. The animals were randomly divided into two groups of eight (same gender distribution) and reared in an environmentally controlled independent nursery.

Water and feed were made available from time to time in the nursery, and feed was used for weaning pills [19% crude protein] without commercially available antimicrobials.

All weaning pigs were euthanized by intravenous injection of sodium barbiturate and then necropsied. Among the 24 weaning pigs, DNA from the stomach and esophagus sections was extracted using a Qiagen Dneasy tissue kit according to the instructions provided at necropsy. 3 μl of extracted DNA was used to test for the presence of Helicobacter spp. In the biopsy tissue sample. Polymerase chain reaction (PCR) was performed twice in each sample, and seven control DNA samples were treated together for each PCR reaction. There was no contradiction between the PCRs performed.

result:

Processing Coding:

Group 1: no treatment (negative control)

Group 2: 0.1 w / v% overactivated polymer antimicrobial agent prepared according to Example 10b; 30mg / kg / day /

Group number Stomach area Esophagus One - - One - + One - + One + - One - - One - + One - - One - + 2 - - 2 - - 2 - - 2 - - 2 - - 2 - - 2 - - 2 - -

PCR results of Helicobacter spp using pre-optimized genus specific primers (+ indicates positive detection of Helicobacter spp.,-Indicates no detection)

In group 1 (no treatment), five positive PCR results were shown for Helicobacter species (1st- gastric, 4-esophageal), while in group 2 (0.1w / v% polymer antimicrobial) no positive PCR results were seen.

conclusion:

Acetal derivatives of poly (2-propenal, 2-propenoic acid) of 0.1w / v% significantly reduced the frequency of porcine Helicobacter spp. (Χ ² : P <0.025) in the stomach and esophageal mucosa of weaning pigs. Let's do it.

Example 20

Comparative Example 20 (a)

This example illustrates a method for preparing poly (2-propeneal, 2-propene acid) that has not been overactivated.

0.8w / w% sodium hydroxide

9.90 kg of deionized water was added to a 10 L stainless steel barrel, and 0.08 kg of sodium hydroxide was added to the water, followed by stirring until dissolved.

polymerization

100.1 kg of deionized water was placed in a 200 L stainless steel keg, and 4.99 kg of 0.8 w / w% sodium hydroxide solution was added to the keg. The solution was equilibrated to 15-20 ° C. At the same time, 20 kg of acrolein monomer was added and the remaining 0.8w / w% sodium hydroxide solution was added to this 200L barrel at a constant rate over 1 hour so that the pH was maintained at 10.5 to 11.0 and the temperature did not rise above 30 ° C. The polymerization was continued for 90 minutes.

wash

The polymerization mixture was filtered / centrifuged and the polymer was washed with deionized water until the pH of the wash water was below 7.0. The approximate yield was 8 kg.

dry

The polymer was dried in air and then heated in an oven in the following order.

step time Temperature One 2 hours 25 ℃ 2 1 hours 40 ℃ 3 1 hours 70 ℃ 4 1 hours 75 ℃ 5 2 hours 85 ℃

Dissolution

400 L of water was added to a 500 L barrel, and then 4 kg of sodium carbonate was added and stirred until dissolved. 8 kg of heated anhydrous polymer was added slowly and stirred for 30 minutes.

The resulting polymer was found to have an approximate solubility of 90-95% in 1w / w% sodium carbonate.

Example 20 (b)

This example illustrates the preparation of an acrolein polymer in which the polymer of Comparative Example 20 (a) is overactivated.

Base manufacturing

0.4 kg of sodium carbonate was dissolved in 30.6 kg of water in a suitable vessel and 64 kg of polyethylene glycol 200 was added to this mixing vessel. Agitation was started with a mechanical stirrer and the PEG200 was heated to 65 ± 3 ° C. To this PEG200 5 kg of the anhydrous acrolein polymer of Example 20 (a) was added and stirred until a homogeneous mixture was obtained.

Note: Solids may not be completely dissolved at this stage.

Thus, sodium carbonate solution is slowly added to the glycol mixture at a rate such that the pH of the solution can be maintained in the range from 3.5 to 9.0.

This solution was stirred at 65 ± 3 ° C. for 45 minutes.

Note: The pH should range from 7 to 9 and the temperature should range from 65 ± 3 ° C.

Transient activation

The mixing vessel was covered and heated to 100 ° C. for 4 hours. The resulting overactivated polymer was observed to be miscible with water in all ratios.

Example 21

In Example 14 of PCT / AU9600328, the poly (2-propenal, 2-propenic acid) polymer in 0.5w / w% sodium carbonate solution demonstrates anticancer activity against Ehrlich ascites cell lines in mouse models. have.

Anticancer Activity of Poly (2-propenal, 2-propene Acid) Polymer [Example 20 (a)] Against Anticancer Activity of Overactivated Polymer [Example 20 (b)]. As an in vitro model of gastrointestinal cancer, human colon cancer cell line HT-29 was used. Poly (2-propeneal, 2-propene acid) was used as a 5w / w% concentrate. Plots were shown and tested for obtaining IC ₅₀ by incubating these cancer cells with various concentrations of polymer.

Way

HT-29 cells (human colon cancer cells) were seeded in wells of 96 well culture plates and incubated overnight at 37 ° C. in a humidified 5% CO ₂ , 95% air atmosphere. The polymer [poly (2-propenal, 2-propenic acid) polymer of Comparative Example 20 (a) and the overactivated polymer of Example 20 (b)] was dissolved in water and concentrated at 10-fold in the 4 log range using medium. Diluted with water. 100 μl of each solution was then added to each of five wells. After incubation of this plate for an additional 72 hours, the viable cell number was assessed by sulforhodamine B assay [Skehan et al., (1990) J. Nat. Cancer Inst. 82: 1107-1112; Monks et al., (1991) J. Nat. Cancer Inst. 83: 757-766. The cells were fixed with 10% cold trichloroacetic acid at 4 ° C. for 1 hour, then the plates were washed with distilled water, left to air dry and then 0.4% sulforhodamine B (product of Aldrich) (v / v) in 1% acetic acid. ) For 30 minutes. Then washed twice with distilled water to remove unbound dye and finally with 1% acetic acid. The protein bound dye was then dissolved in 10 mM unbuffered Tris base and the absorbance was read at 550 nm using an autoplate reader. Average absorbance of each drug dose is expressed as the percentage of control untreated well absorbance.

The test results are shown in Table 24.

The poly (2-propeneal, 2-propene acid) polymer of Comparative Example 20 (a) exhibited an average IC ₅₀ during two tests at 0.030%; Poly (2-propeneal, 2-propene acid) polymer was designated as 100%. This translates to 0.0015 w / w% of the active polymer.

The overactivated polymer of Example 20 (b) exhibited an average IC ₅₀ during four 0.025% tests. This translates to 0.00125 w / w% of the overactivated polymer and suggests that the overactivated polymer has strong anticancer activity.

Test compound Drug Exposure Time (hr) IC ₅₀ (%) Average IC ₅₀ (w / w%) IC ₅₀ average (w / w% of active polymer) Comparative Example 20 (a) 72 0.037, 0.023 0.030 0.0015 Example 20 (b) 72 0.017, 0.034, 0.025, 0.023 0.025 0.00125

IC ₅₀ of CHEMEQ ^RTM Polymer Antimicrobial Agents for HT-29 Human Colon Cancer Cells

IC ₅₀ is the concentration required to inhibit cell proliferation 50%.

Finally, it will be apparent that various other modifications and / or modifications may be made without departing from the spirit of the invention as outlined herein.

Claims (32)

Derivatives of poly (2-propenal, 2-propenoic acid) formed by reacting an organic compound containing at least one hydroxyl group with poly (2-propenal, 2-propenoic acid) to form a protected carbonyl group A method of treating or preventing gastrointestinal disease in an animal (including humans), comprising administering to the animal an effective amount of a polymer. The method of claim 1, wherein the polymer is administered orally. The method of claim 1, wherein the animal is suffering from gastroenteritis, ulcers, diarrhea and at least one gastrointestinal disease selected from the group consisting of gastrointestinal cancer and insufficient weight gain of heterogeneous triggers. The method of claim 1, wherein the animal is suffering from at least one of diarrhea, gastroenteritis, and dysentery. The method of claim 1, wherein the animal is selected from the group consisting of dogs, pigs, sheep, horses, cows, cats, poultry, ducks, turkeys and quails. The method of claim 1, wherein the animal is selected from ruminants and the polymer is rectally administered. The method of claim 1, wherein the animal is selected from poultry and pigs. The method of claim 1, wherein the animal is a partially grown pig. A method of treating or preventing E. coli in weaning pigs, including orally administering an antimicrobial effective amount of the polymer of claim 1 to a young pig after weaning. The method of claim 1, wherein the antimicrobial agent of claim 1 is administered at a dosage of 0.05 to 5000 mg / kg / day. The method of claim 1, wherein the antimicrobial agent of claim 1 is administered at a dosage ranging from 0.5 to 500 mg / kg / day. The method of claim 1, wherein the pig is administered an antimicrobial dosage in the range of 0.05-50 mg / kg / day. The method of claim 1 wherein the gastrointestinal disease is E. coli, Salmonella, P. ah rugi labor (P.aeruginosa), Helicobacter pylori (Helicobacter), Proteus (Proteus), Enterobacter bacteria (Enterobacteria), yeast (Yeast), protozoa (Protozoa ), Clostridia (Clostridia), Shigella (Shigella) and koksi Guardia (the characteristic that the method Coccidia) is one or more microorganisms is selected from the group consisting of a cause. Poly (2-propenal) formed by reacting an organic compound containing a hydroxyl group selected from alkanols, phenols, polyols and mixtures thereof with poly (2-propene, 2-propenic acid) to form a protected carbonyl group A method of treating or preventing diseases of the gastrointestinal tract caused by Helicobacter infection, comprising administering to the stomach a therapeutic amount of a polymer comprising a derivative of 2-propene acid). The method of claim 1, wherein the gastrointestinal disease is caused by at least one of enterotoxin E. coli and β-hemolytic E. coli. A method of treating or preventing necrotic enteritis in poultry, comprising administering to the poultry an effective amount of the antimicrobial agent of claim 1. The method of claim 1, wherein the antimicrobial agent is administered with an additional chemotherapeutic agent, wherein the additional chemotherapeutic agent is adsorbed onto the antimicrobial agent to reduce membrane penetration. A method of treating or preventing poultry coccidiosis comprising administering to the poultry an antimicrobial effective amount of the antimicrobial agent of claim 1. The method of claim 1 wherein the derivative comprises a plurality of protected carbonyl groups selected from at least one hemiacetal group and an acetal group. The method of claim 20, wherein the protected carbonyl group comprises an acetal group. The method of claim 1 wherein the alcohol is selected from alkanols, phenols, polyols and mixtures thereof. The method of claim 22, wherein the alcohol is selected from at least one polyol. The method of claim 23, wherein the polyol comprises polyalkylene glycol. The method of claim 23, wherein the polyol comprises polyethylene glycol. 24. The method of claim 23, wherein the polyol is polyethylene glycol having a molecular weight between 200 and 2000. Animals and derivatives of poly (2-propeneal, 2-propenoic acid) formed by reacting an organic compound containing at least one hydroxyl group with poly (2-propaneal, 2-propene acid) to form a protected carbonyl group An antimicrobial agent for treating or preventing gastrointestinal diseases in an animal by administering to the gastrointestinal antimicrobial composition comprising a pharmaceutically or veterinary acceptable inert carrier for gastrointestinal administration. The carrier of claim 27 wherein the carrier for gastrointestinal administration is water, controlled release polymers, olive oil, peanut oil, sesame oil, sunflower oil, arachis oil, coconut oil, liquid paraffin, ethylene glycol, propylene glycol, polyethylene glycol, ethanol, An antimicrobial agent for treating or preventing gastrointestinal diseases, characterized in that it is selected from the group consisting of propanol, isopropanol, glycerol, fatty acid alcohols, triglycerides, polyvinyl alcohols, partially hydrolyzed polyvinyl acetates and mixtures thereof. The antimicrobial agent according to claim 27, which is in the form of a feed additive or a drinking water additive containing 0.1 to 70% by weight of the antimicrobial agent according to claim 1. A feed or drinking water composition of an animal (including human) containing an antimicrobial effective amount of the antimicrobial agent according to claim 1 and a feed or water. 31. The feed composition of the animal according to claim 30, wherein the antimicrobial agent is present in an amount of 0.001 to 25% by weight based on the total feed or drinking water composition. An antimicrobial composition comprising the antimicrobial agent according to claim 1 and an additional active agent selected from the group consisting of an additional antimicrobial agent and a chemotherapeutic agent. The method of claim 31, wherein the additional antimicrobial agent is based on the weight of the composition: (a) phenol in an amount of 0.1 to 10%; (b) isothiazolinone in an amount of 0.001 to 1%; (c) alkyl parabens in an amount of 0.02 to 2% and (d) lower alkanols in an amount of 20 to 99.9% Antimicrobial composition characterized in that it comprises at least one of.