WO2009138455A1

WO2009138455A1 - Enzyme composition for reducing the release of pharmaceutical active ingredients into the environment

Info

Publication number: WO2009138455A1
Application number: PCT/EP2009/055834
Authority: WO
Inventors: Giuseppe Primiceri; Elena Roda; Alessandra Stefan; Lucio Panizza; Alejandro Hochkoeppler
Original assignee: Archimede R&D S.R.L.; Bioikos Ambiente S.R.L.
Priority date: 2008-05-16
Filing date: 2009-05-14
Publication date: 2009-11-19
Also published as: ITBO20080297A1

Abstract

A composition comprising one or more enzymes selected from the group consisting of oxidoreductases, transferases, hydrolases, lyases, isomerases and ligases for reducing the release into the environment of one or more pharmaceutical active ingredients, derivatives and metabolites thereof by a subject undergoing pharmacological therapy with the one or more pharmaceutical active ingredients, by means of an enzyme reaction of the one or more pharmaceutical active ingredients, derivatives and metabolites thereof with the one or more enzymes, the composition also comprising the one or more pharmaceutical active ingredients or being administered simultaneously, sequentially or separately with respect to the one or more pharmaceutical active ingredients.

Description

ENZYME COMPOSITION FOR REDUCING THE RELEASE OF PHARMACEUTICAL ACTIVE INGREDIENTS INTO THE ENVIRONMENT Technical Field The present invention relates to an enzyme composition for reducing the release into the environment of pharmaceutical active ingredients administered to a subject, so as to make the environmental impact of such active ingredients return within acceptable limits. The composition is used both in the human field and in the veterinary field. Background Art

The European standards that regulate the marketing of pharmaceutical drugs, both for veterinary use and for human use, require the applicant for the AIC (Autorizzazione all'Immissione in Commercio, Marketing Authorization) to submit an assessment of the potential environmental risk linked to the use of the medications and to their excretion into the environment by the subjects to which these medications are administered. The risk estimate must take into account not only the molecule or molecules constituting the active ingredient of the medication but also any products of degradation that might form during metabolization. Moreover, the environmental impact caused by use of the pharmaceutical drug must be assessed both in soil and in water, depending on the type of use of such drug.

The assessment of the risk that arises from introducing into the environment a pharmaceutical active ingredient can refer to indexes such as PECsoiL ("Predicted Environmental Concentration Soil"), PECSURFΛCEWATER (Predicted Environmental Concentration for Surface Water) and EICAQUATIC ("Environmental Introduction Concentration Aquatic"). These indices are theoretical values, calculated by means of equations, for pharmaceutical drugs that will be excreted into the ground, into the water and will be used by human beings. The theoretical limit values, which are identical for all drugs, are equal to 100 μg/kg, 1 μg/1 and 0.01 μg/1 respectively for PECSOIL, PECsuRFACEWATER and EICAQUATIC, and represent the values which, if exceeded, make it mandatory to submit specific studies aimed at demonstrate the degree of safety of the pharmaceutical drugs or metabolites thereof that are dispersed into the environment. Among the reference guidelines, mention can be made of EMEA/CHMP/SWP/4447/00 and EMEA/CVMP/ERA/418282/2005. These analyses of risk assessment are necessary in order to determine the possible impact of the release into the environment of substances that have a pharmacological activity and might influence the ecosystem into which they are introduced. For example, the release of active ingredients that have an antibiotic action can cause, in the microorganisms that are present in the environment, the onset of antibiotic resistance phenomena, with negative effects on the therapeutic capacity of the antibiotic itself. The need to perform in-depth studies on the environmental impact of pharmaceutical drugs therefore entails high costs. In the veterinary field, these costs are further worsened by the need for disposal as special waste of the excretions of farm animals subjected to pharmacological treatments. The presence of pharmacological substances in such excretions in fact does not allow their release into the environment and makes it necessary to perform expensive disposal treatments.

There is, therefore, the need for a method for reducing the environmental impact of pharmaceutical active ingredients released by a subject to which they have been administered, which allows to obviate partly or fully the need to perform expensive and time-consuming environmental risk assessment analyses and allows to reclassify as non- dangerous waste the excretions of the subject (a farm animal or a human being) undergoing pharmacological therapies. Disclosure of the Invention The aim of the present invention is to provide an enzyme composition that allows to reduce the environmental impact of pharmaceutical active ingredients released into the environment by a subject to which they have been administered.

Within this aim, an object of the invention is to provide an enzyme composition that reduces the environmental impact of pharmaceutical active ingredients prior to their actual release into the environment by the subject to which they have been administered.

Another object of the invention is to provide an enzyme composition that reduces the environmental impact of a pharmaceutical drug and allows to reduce or eliminate the need for disposal operations on the excretions of the subject to which pharmaceutical active ingredients have been administered.

Another object of the present invention is to provide an enzyme composition that reduces the quantity of pharmaceutical active ingredients released into the environment by the subjects to which they have been administered.

Another object of the invention is to provide an enzyme composition for reducing the environmental impact of pharmaceutical active ingredients that is highly reliable, relatively easy to provide and at low costs. This aim and these and other objects, which will become better apparent hereinafter, are achieved by a composition comprising one or more enzymes selected from the group consisting of oxidoreductases, transferases, hydrolases, lyases, isomerases and ligases for reducing the release into the environment of one or more pharmaceutical active ingredients, derivatives and metabolites thereof by a subject undergoing pharmacological therapy with the one or more pharmaceutical active ingredients, by means of an enzyme reaction of the one or more pharmaceutical active ingredients, derivatives and metabolites thereof with the one or more enzymes, said composition also comprising the one or more pharmaceutical active ingredients or being administered simultaneously, sequentially or separately with respect to the one or more pharmaceutical active ingredients.

Brief description of the drawings

Further characteristics and advantages of the invention will become better apparent from the following detailed description and from the accompanying drawings, wherein:

Figure 1 is a chart of the calibration curve obtained by measuring the absorbance of standard samples provided with an ELISA (Enzyme-Linked Immunosorbent Assay) kit used to assess the activity of the enzymes indicated in Table 1 on the sulfonamide antibiotic sulfadiazine;

Figure 2 is a chart of the absorbance values determined by ELISA assay on samples of sulfadiazine incubated with the enzymes indicated in Table 1 at pH 4.5 (shaded histograms) and pH 7 (white histograms);

Figure 3 is a chart of the absorbance values determined by means of an ELISA assay on samples of sulfadiazine incubated respectively in the absence of an enzyme, with porcine liver esterase and S. cerevisiae esterase. Ways of carrying out the Invention

The composition herein described comprises one or more enzymes whose catalytic activity allows them to favor chemical reactions at the level of the active ingredients contained in the pharmaceutical products administered to a subject being treated. These chemical reactions can degrade the active ingredient or convert it into a substance whose environmental impact is minimal with respect to the active ingredient. For example, conversion can consist in functional izing the active ingredient with particular functional groups, such as to simplify disposal and reduce its retention time in the environment.

Of course, the activity of the one or more enzymes of the composition according to the invention can also occur so as to affect any derivatives or metabolites of the active ingredients that form during the cycle of pharmacological activity and are dangerous for the environment as much as the active ingredient from which they derive.

According to the invention, the composition comprising the one or more enzymes can be administered to the subject simultaneously, sequentially or separately with respect to the one or more active ingredients or the enzyme composition can comprise the one or more active ingredients.

In a preferred embodiment, the composition according to the invention further comprises the one or more active ingredients. This embodiment is particularly advantageous, since it allows to administer to the subject both the pharmaceutical active ingredient and the one or more enzymes to reduce its environmental impact by resorting to a single formulation.

In an alternative embodiment, the composition according to the invention can comprise the one or more enzymes in a quantity from 0.001% to 50% of the quantity of the one or more active ingredients administered to the subject, which corresponds to an enzyme:active ingredient ratio between 0.00001 : 1 and 0.5: 1. The quantity of the one or more enzymes can be equal for example to 5%, which corresponds to an enzyme:active ingredient ratio of 0.05: l .

According to the invention, the composition can be administered to the subject orally. As an alternative, administration can occur parenterally, for example by injection. Preferably, administration is oral.

In a preferred embodiment, the reaction of the one or more pharmaceutical active ingredients, derivatives and metabolites thereof with the one or more enzymes of the composition occurs at the level of the intestine of the subject.

Without intending to be constrained to any theory in particular, it is believed that the enzymes, as a consequence of the administration of the composition described here, become located at the level of the intestine of the subject, while the pharmaceutical active ingredients are absorbed and diffused by the circulatory system of the subject to the tissues. During their intestinal permanence, the one or more enzymes come into contact with the molecules of the pharmaceutical active ingredients, their derivatives or metabolites sent to the excretion system at the end of their cycle of pharmacological activity. Therefore, the chemical reactions catalyzed by the one or more enzymes and adapted to degrade the active ingredients or to convert them into substances that are less toxic or not at all toxic for the environment occur in the intestine.

Preferably, the one or more enzymes of the composition according to the invention have an enzyme activity at a pH higher than 7 that is 10%, more preferably 20%, higher than enzyme activity at a pH lower than 5. The higher activity at substantially basic pH values allows the one or more enzymes to catalyze efficiently the degradation of the one or more active ingredients only in body environments that corresponds to specific conditions in terms of pH, thus avoiding an early and unwanted enzyme reaction. For example, this allows the one or more enzymes to degrade or modify efficiently the one or more active ingredients at the intestinal level, where the pH is higher than 7, rather than at the gastric level, where the pH is acid and markedly lower than 5. Advantageously, in this manner it is possible to avoid a degradation of the one or more active ingredients prior to their absorption.

In an alternative embodiment, the composition according to the invention can be formulated in a form of dosage that has a gastroresistant coating. The expression "gastroresistant coating" is used to reference a protective film that is applied to the form of dosage and whose surface is stable at the acid pH that characterizes the stomach, breaking up instead in basic pH conditions such as the ones that can be observed at the intestinal level. The gastroresistant coating therefore prevents early release of the content of the form of dosage, further protecting it against conditions that are potentially harmful for its functionality. Generally, substances sensitive to the basic conditions of the intestinal environment, characterized for example by the presence of ester groups that are hydrolyzed in the intestine but not in the stomach, or certain waxes, again of the ester type, are used for gastroresistant coatings.

In another embodiment, the one or more enzymes of the composition according to the invention can be enclosed in a polymeric matrix for controlled release. In this manner it is possible to control the times and quantities of the release of the one or more enzymes. The expression "polymeric matrix for controlled release" is used to reference a coating made of polymeric material that is formulated to dissolve slowly and release over time the active ingredient covered by it. For example, the polymeric material can be an acrylic polymer, a cellulose derivative or a synthetic biodegradable polymer such as polylactic acid (PLA), polyglycolic acid or polylactic-co-glycolic acid (PLGA).

The enzymes used in the composition according to the invention are selected from the group consisting of oxidoreductases, transferases, hydrolases, lyases, isomerases and ligases. More preferably, the enzymes are selected from the group consisting of oxidoreductases and hydrolases.

The hydrolases in turn can be selected among proteases, lipases and esterases. The oxidoreductases can be for example laccases or tyrosinases. In a particularly preferred embodiment, the one or more enzymes can be selected from the group consisting of porcine esterase, yeast esterase, and enzymes derived from snail acetone powder.

The expression "enzymes derived from snail acetone powder" is used to reference an enzyme complex obtained from snail tissues by extraction with acetone.

The pharmaceutical active ingredients that constitute the target of the activity of the enzymes of the composition according to the invention and whose environmental impact is sought to be reduced can be characterized by the presence of one or more chemical functionalities within their structure. These functionalities can be for example hydroxyl groups (-OH), carbonyl groups (>C=O), carboxyl groups (-COOR), a halogen (-Cl, -Br, -I, -F), a thiol group (-SH), an ether group (R-O-R), an amine group (-NR₃), a cyano group (-CN), a nitroxide group (-N=O), an oxime (-CN0H), an anhydride group (-R-C(=O)-O-(C=O)-R-), a -C=S- group, a -C=N- group, a phosphate group (-PO₄), a nitrate group (-NO₃), a nitrite group (-NO₂), a sulfate group (-SO₄), a -C=C- or -N=N- double bond and a -C≡C- or -N≡N- triple bond, where each R, independently of each other, can be hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heterocycloalkyl or heteroaryl.

The term "alkyl" identifies a linear or ramified hydrocarbon chain, which contains at least one carbon atom. Non-limiting examples of an alkyl are represented by a functional group selected among methyl, ethyl, n- propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylbutyl, 3-methylhexyl, 3,3-dimethylbutyl, 2,2- dimethylpentyl, n-octyl, n-nonyl and n-decyl. The term "alkenyl" identifies a linear or ramified hydrocarbon chain that contains at least two carbon atoms and at least one carbon-carbon double bond formed by the removal of two hydrogen atoms. Non-limiting examples of an alkenyl are represented by a functional group selected among ethenyl, 2-propenyl, 2-methyl-2-propenyl, 3-butenyl, 4-pentenyl, 5- hexenyl, 2-heptenyl, 2-methyl-l-heptenyl and 3-decenyl.

The term "alkynyl" identifies a linear or ramified hydrocarbon chain that contains at least two carbon atoms and at least one carbon-carbon triple bond. Non-limiting examples of an alkynyl are represented by a functional group selected among 1-propynyl, 2-propynyl, 3-butynyl, 2-pentinyl, and 1 - butynyl.

The term "cycloalkyl" identifies a cyclic ring system exemplified by a saturated cyclic hydrocarbon that has 3 to 8 carbon atoms. Non-limiting examples of saturated cyclic systems include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl. The term "aryl" identifies an (aromatic) unsaturated cyclic ring system, which has 3 to 10 carbon atoms. Non-limiting examples of aromatic cyclic systems include the functional groups phenyl and naphthyl.

The term "heterocycloalkyl" identifies a cycloalkyl group as described above, in which the ring contains at least one heteroatom selected independently from the group consisting of nitrogen, oxygen and sulphur. Non-limiting examples of heterocycloalkyl groups include tetrahydropyiTol, tetrahydrofuran, tetrahydrothiophene, piperidine, tetrahydropyrane, piperazine and dioxane.

The term "heteroaryl" identifies an aryl group as described above, in which the ring contains at least one heteroatom selected independently from the group consisting of nitrogen, oxygen and sulphur. Non-limiting examples of heteroaryl groups include benzoimidazol, benzofuran, benzothiazol, benzothiophene, imidazole, indazole, pyridine, pyrrole, pyrimidine or triazole. Preferably, the one or more active ingredients can be antibiotics, antiparasitic agents, chemotherapy agents, anti-inflammatory or analgesic agents. For example, the antibiotics can be tetracyclines. The chemotherapy agents can be for example sulfonamides.

The composition according to the invention can be used to reduce the environmental impact of pharmaceutical active ingredients released by subjects such as human beings or animals. In a preferred embodiment of the present invention, the subject is a human being. In an alternative embodiment, the subject is an animal, in particular a farm animal selected from the group consisting of bovines, sheep, goats, equines, swine, poultry, rabbits and fish.

In another aspect, the present invention also relates to a method for reducing the release into the environment of one or more pharmaceutical active ingredients, derivatives and metabolites thereof by a subject undergoing pharmacological therapy with the one or more pharmaceutical active ingredients, by means of an enzyme reaction of the one or more pharmaceutical active ingredients, derivatives and metabolites thereof with a composition comprising one or more enzymes selected from the group consisting of oxidoreductases, transferases, hydrolases, lyases, isomerases and ligases, said composition comprising also the one or more pharmaceutical active ingredients, or being administered simultaneously, sequentially or separately with respect to the one or more pharmaceutical active ingredients. Example 1 The degradation effectiveness of 16 different enzymes, listed in Table 1 , with respect to the sulfonamide antibiotic sulfadiazine was evaluated. An enzyme immunoassay (ELISA) specific for detection of sulfadiazine and its metabolites was used for this analysis. In particular, a kit manufactured by Euro-Diagnostica (Arnhem, Netherlands) was employed. Sulfadiazine is a sulfonamide used extensively for therapeutic and prophylactic purposes in veterinary medicine, so much that in many countries, such as Europe, the United States and Canada, a maximum residual value (known as MRL, maximum residue limit) of sulfonamides in the tissues of animals intended for human food consumption that corresponds to 100 μg/kg has been established. The ELISA kit used here reveals up to 0.1 ng/ml of sulfadiazine (the reading range is 0.1-10 ng/ml). In particular, the ELISA assay used here is of the competitive type using polyclonal rabbit antibodies that are specific for recognition of any sulfadiazine that might be present in the sample being analyzed. Competition occurs between the "free" sulfadiazine of the sample and the sulfadiazine added in the assay. The latter is conjugated with an enzyme capable of hydrolyzing a chromogenic substrate, producing a coloured compound (yellow) with an absorption maximum at 450 nm. The intensity of the colour that is developed (which can be measured in a spectrophotometer) is inversely proportional to the concentration of sulfadiazine that is present in the sample. The procedure followed to assess the enzyme efficiency of sulfadiazine degradation entails various steps:

- adding the sample in a well of the ELISA plate;

- adding the enzyme-sulfadiazine conjugate; - adding the anti-sulfadiazine antibody;

— incubating for 1 hour at 4° C;

— washing with a suitable buffer (3 times);

- adding the substrate for the enzyme;

— incubating for 30 minutes; - adding a solution of sulphuric acid to facilitate the development of the colour;

— measuring absorbance at 450 nm;

- processing and interpreting the data.

Alongside the unknown specimens, 6 different standard samples were analyzed which contained a known concentration of sulfadiazine: 10 ng/ml, 5 ng/ml, 1 ng/ml, 0.5 ng/ml, 0.1 ng/ml and 0.05 ng/ml. In this manner it was possible to obtain a calibration curve to be used to convert the absorbance values into sulfadiazine concentration. White absorbance (sample containing only the buffer) was subtracted from all the absorbance values. The samples analyzed by ELISA assay were obtained by incubation of sulfadiazine in the presence of each one of the enzymes listed in Table 1.

Table 1

# , Enzyme # Enzyme

1 I Pancreatic chymotrypsin 9 , Liver acetone powder

2 i Porcine liver esterase 10 Papain

3 R. oryzae esterase 11 A. oryzae protease

4 S. cerevisiae Esterase ₍ i 12 S. griseus protease # I Enzyme # Enzyme

5 R. vernicifera laccase 13 Snail acetone powdei

6 ' T. versicola laccase 14 Aerobacter sulfatase

7 C. rugosa lipase 15 Patella sulfatase

8 Porcine pancreatic lipase 16 Fungal tyrosinase

Each enzyme was dissolved in water so as to obtain a 10 mg/ml solution. The sulfadiazine was instead dissolved in 200 mM NaOH so as to prepare a 100 mM solution. For each enzyme, incubation was performed at pH 4.5, using a 50 mM phosphate buffer and at pH 7, using a 500 mM

MOPS buffer.

The samples used therefore consisted of 800 μl of enzyme solution (10 mg/ml), 100 μl of sulfadiazine (100 mM) and 100 μl of buffer. The samples were incubated at 37°C overnight. The next day, 50 μl of each reaction were used to perform the ELISA assay.

The calibration curve was determined by using the sulfadiazine standards included in the ELISA kit, so as to obtain the straight line shown in Figure 1 , from which it is possible to determine the unknown concentration of sulfadiazine in the analyzed samples and determine enzyme activity.

The enzyme activity of the individual enzymes on sulfadiazine was determined in triplicate by measuring the absorbance of the samples incubated with the 16 enzymes at pH 4.5 and pH 7. The results obtained are plotted in chart form in Figure 2, where the enzymes are identified by means of the numbering given in Table 1, the shaded histograms refer to the absorbance measured for the reaction performed at pH 4.5, and the white histograms refer to the absorbance for the reaction performed at pH 7. As can be seen, all the enzymes exhibit activity against sulfadiazine. In particular, porcine liver esterase and S¹. cerevisiae esterase have shown considerable activity on sulfadiazine and therefore additional determinations of the activity of these enzymes were conducted.

Example 2

Porcine liver esterase and S. cerevisiae esterase were used to repeat in triplicate the absorbance measurements on samples subjected to the ELISA test with the methods described previously in example 1.

As can be seen from the chart in Figure 3, the activity of both enzymes is evident from the comparison of the absorbance detected in the solutions containing sulfadiazine and the enzymes (2 = S. cerevisiae esterase, 3 = porcine liver esterase) with respect to the absorbance of the solution that contains only sulfadiazine (1 = negative control).

Example 3

A spectrophotometric assessment of the degree of enzyme degradation performed by porcine liver esterase and S. cerevisiae esterase on sulfadiazine was performed, assessing the absorbance variation at 255 nm.

The samples were prepared by using 5 μl of a solution of 10 rnM sulfadiazine in 200 mM NaOH, 100 μl of an aqueous solution of enzyme

(porcine liver esterase or S. cerevisiae esterase) at a concentration of 1 mg/ ml, 395 μl of water and 500 μl of a 100 mM phosphate buffer at pH 7. 100 μl of water were used for the control sample instead of the enzyme solution.

The absorbance of the samples at 255 nm was then measured in triplicate both at time 0 (pre-incubation), and after 20 hours at 37°C, subsequently calculating the mean and the standard deviation and subtracting from the absorbance values of the reactions the measurements obtained only for the enzyme mixtures without sulfadiazine. The results of the measurements of the absorbance and the percentages of enzyme degradation are listed in Table 2, where sample 1 corresponds to the porcine liver esterase enzyme and sample 2 corresponds to the S. cerevisiae esterase enzyme. Table 2 Sample T = Oh '^■ T = 2Oh _| Degradation

Absorbance i Std. Dev. Absorbance Std. Dev.

Control 1.372 0.058 1.370 0.041 0%

1.417 0.008 1.026 0.020 27.59%

1.384 0.028 1.165 0.010 15.82%

As can be seen, porcine liver esterase allows to achieve a sulfadiazine degradation of 27.59%. In practice it has been found that the composition according to the invention fully achieves the intended aim, since it allows to reduce the environmental impact of a pharmaceutical product released into the environment by a subject to which the product has been administered by enzyme reaction of the pharmaceutical active ingredient with the one or more enzymes of the composition.

It has also been observed that the composition according to the invention reduces the impact on the environment caused by the excretions of subjects undergoing pharmacological therapy, obviating or otherwise reducing the need for disposal as special waste of such excretions, particularly in the field of animal farms.

Moreover, it has also been observed that the composition according to the invention, by reducing the introduction of pharmaceutical active ingredients into the environment, also leads to a simultaneous reduction of the dangers caused by the diffusion into the environment of such pharmaceutical substances, such as allergic phenomena or resistance to antibiotics on the part of microorganisms.

The composition thus conceived is susceptible of numerous modifications and variations, all of which are within the scope of the appended claims; all the details may further be replaced with other technically equivalent elements.

In practice, the materials used, as well as the dimensions, may be any according to requirements and to the state of the art.

The disclosures in Italian Patent Application No. BO2008A000297 from which this application claims priority are incorporated herein by reference.

Where technical features mentioned in any claim are followed by reference signs, those reference signs have been included for the sole purpose of increasing the intelligibility of the claims and accordingly, such reference signs do not have any limiting effect on the interpretation of each element identified by way of example by such reference signs.

Claims

1. A composition comprising one or more enzymes selected from the group consisting of oxidoreductases, transferases, hydrolases, lyases, isomerases and ligases for reducing the release into the environment of one or more pharmaceutical active ingredients, derivatives and metabolites thereof by a subject undergoing pharmacological therapy with the one or more pharmaceutical active ingredients, by means of an enzyme reaction of the one or more pharmaceutical active ingredients, derivatives and metabolites thereof with the one or more enzymes, said composition also comprising the one or more pharmaceutical active ingredients or being administered simultaneously, sequentially or separately with respect to the one or more pharmaceutical active ingredients.

2. The composition according to claim 1 , wherein the reaction of the one or more pharmaceutical active ingredients, derivatives and metabolites thereof with the one or more enzymes occurs at the level of the intestine of the subject.

3. The composition according to claim 1 or 2, wherein the one or more enzymes have an enzyme activity, at a pH of more than 7, that is 10% higher than the enzyme activity at a pH lower than 5.

4. The composition according to one or more of claims 1 to 3, wherein the one or more enzymes are selected from the group consisting of porcine esterase, yeast esterase and enzymes derived from snail acetone powder.

5. The composition according to one or more of claims 1 to 4, wherein the one or more enzymes are in a quantity that corresponds to an enzyme:active ingredient ratio between 0.00001 : 1 and 0.5 : 1.

6. The composition according to claim 5, wherein the enzyme: active ingredient ratio is 0.05: 1.

7. The composition according to one or more of claims 1 to 6, wherein the one or more active ingredients comprise one or more chemical functionalities selected from the group consisting of hydroxy 1, carbonyl, -COOR, -Cl, -Br, -I₅ -F, -SH, R-O-R, -NR₃, -CN, -N=O, -CNOH, -R-C(=Q)- 0-(C=O)-R-, -C=S-, -C=N-, -PO₄, -NO₃, -NO₂, -SO₄, -C=C-, -N=N-, -C≡C-, - N≡N-, where each R, independently of each other, is hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heterocycloalkyl or heteroaryl.

8. The composition according to one or more of claims 1 to 7, wherein the one or more active ingredients are selected from the group consisting of antibiotics, antiparasitic agents, chemotherapy agents, antiinflammatory agents and analgesic agents.

9. The composition according to one or more of claims 1 to 8, further comprising the one or more pharmaceutical active ingredients.

10. The composition according to one or more of claims 1 to 9, characterized in that it is administered orally or parenterally.

11. The composition according to claim 10, wherein administration is oral.

12. The composition according to one or more of claims 1 to 1 1 , characterized in that it is formulated in a form of dosage that has a gastroresistant coating.

13. The composition according to one or more of claims 1 to 12, wherein the one or more enzymes are enclosed in a polymeric matrix with controlled release.

14. The composition according to one or more of claims 1 to 13, wherein the subject is a human being.

15. The composition according to one or more of claims 1 to 13, wherein the subject is an animal selected from the group consisting of bovines, sheep, goats, equines, swine, poultry, rabbits and fish.