WO2009030205A2 - Novel gelatine-active ingredient conjugates - Google Patents

Abstract

The invention relates to a pharmaceutical and/or cosmetic composition containing a conjugate of a natural or synthetic active ingredient comprising a reactive group or several reactive groups, and gelatine.

Description

 New gelatine-drug conjugates.

Field of the invention

The invention relates to a pharmaceutical composition containing a natural or synthetic active ingredient, the use of such a pharmaceutical composition for the manufacture of a medicament for the treatment of a disease which has changed

Metabolism of diseased cells is associated, a method for producing such a pharmaceutical composition and a method of treatment using such a drug.

Prior art and background of the invention.

More than 3000 years ago, essential oils were extracted from plants and / or their flowers in Egypt. The

Users have already been aware of the preservative properties of these natural products, and using these oils, powders, ointments and dressings have been prepared, and others. used in the embalming of dead.

This knowledge of the antibiotic properties of these plant extracts has been preserved from oblivion and expanded by ancient authors such as Dioscurides and Hippocrates, as well as modern physicians such as Avicennia and Culpeper. This knowledge was used in the past century by Gattefosse for the development of "aromatherapy" (D. Krämer, Neue Therapien mit essential oils and precious stones, Isotrop-Verlag, Bad Camberg, ISBN 978-3-940395-00-9).

Despite their known antiseptic properties, the medical applications of these essential oils have been steadily reduced thanks to an increasingly powerful pharmaceutical industry and its constant evolution. Increasingly frequent resistance of germs to modern antibiotics, however, gives reason to develop new applications of "old remedies". However, essential oils in their pure form are particularly distinguished by their toxic effects on the central nervous system, the kidneys and the respiratory tract. In terms of their potency on humans exist - significant differences. Very toxic are e.g. Camphor, eucalyptus (cineole) and peppermint oil

(Menthol). Slightly less toxic are turpentine oil, orange / lemon peel, tea tree and clove oil. In principle, essential oils should only be used in dilute form. At high doses, allergic reactions and asthmatic seizures are possible. The contact of essential oils with eyes or mucous membranes leads to severe irritation.

Regardless of this, cell toxicity is also present in modern synthetic drugs; in some cases, these drugs are even based on them, for example in the

Area of chemotherapy for cancer. This raises the problem of "targeting", ie the active substances only have their effect when they are taken up in diseased cells, wherein the active ingredient must be appropriately masked in such a way that healthy cells the drug on the other hand not or record only reduced. Technical problem of the invention

The invention is therefore based on the technical problem of specifying a pharmaceutical composition which modifies a toxic natural or synthetic active ingredient so that it first develops its toxic effect in the target tissue or cells.

Broad features of the invention and preferred embodiments.

To solve this technical problem, the invention teaches a pharmaceutical and / or cosmetic composition comprising a conjugate of a natural or synthetic active ingredient having a reactive group or having multiple reactive groups with gelatin.

The invention achieves the following. While low molecular weight drugs (Mw 30 to 5000 Da, usually up to 200 Da), for example, low molecular weight, lipophilic essential oils are absorbed very quickly via normal cells of the skin, mucous membranes or inner surfaces such as intestinal wall, etc., this behavior is covalent to gelatin bound low molecular weight drugs not to. Thus, unwanted side effects, such as skin irritation, allergic reactions, etc., are not expected. After a covalent binding of the active ingredient, for example of the essential oil, to gelatin, its biological properties are masked by those of the carrier protein and simultaneously deactivated temporarily. The conjugate does not differ in its biokinetics from the original gelatin. By doing that, normal cells do not absorb proteins or protein fragments to cover their energy needs, healthy organs are not burdened by the uptake of protein-bound drugs. In contrast to healthy tissue (cells) take morbidly altered cells, such as cells in inflammatory processes, strongly proliferating cells z. As cancer cells, cells of atopic dermatitis, etc., gelatin fragments to cover their nitrogen and energy needs. The activation of the drug is therefore only within the affected cells during the lysomal degradation of the

Protein fragment. The active ingredient is released and can locally its effect, such as its cytotoxic effect unfold. Through this process, one is able to introduce active ingredients into the desired cells via a nonspecific process. In contrast, cells with normal metabolism do not

Drug introduction. or recording. The invention finally allows a so-called "targeted therapy" of target tissues.

Finally, the invention makes it possible to make substances which are sparingly soluble in water soluble in water by covalent coupling with the gelatin and thus to considerably improve the pharmacokinetics or absorption, as well as to simplify the technical processing. Finally, gelatin-bound substances are much more stable to oxidative influences when exposed to air, i.e., exposed to air. Shelf life is improved.

A conjugate is a molecule which contains at least one gelatin molecule and one active ingredient molecule in covalent bond. Gelatin is a biopolymer, a peptide, and contains essentially as monomers the amino acids alanine, arginine, aspartic acid, glutamic acid, glycine, histidine, hydroxylysine, hydroxyproline, isoleucine, leucine, lysine,

Methionine, phenylalanine, proline, serine, threonine, tyrasin (type A, not type B), and valine. The amino acid distribution for type A and type B gelatin is different. Main constituents in both cases are glycine (about 25-35 mol%), proline (about 13-18 mol%),

Hydroxyproline (about 13-18 mol%), and alanine (about 15 mol%). The side groups of the amino acids of the polymer partially carry chemical groups, which may be reactive. The average molecular weight of the peptide molecules in gelatin is typically in the range of Mw = 25,000-40,000 Da, in particular Mw = 30,000 to 35,000.

A reactive group of an active agent is a chemical group capable of covalent bonding with a chemical group of a side group of a gelatin peptide. These include u.a. Keto groups, aldehyde groups, hydroxyl groups, carboxyl groups, and amine groups. Essential oils contain especially keto and / or aldehyde groups. Eligible drugs typically have a molecular weight of less than 5,000 Da, more preferably less than 1,000 Da, preferably less than 500 Da, typically less than 200 Da.

The molar ratio of gelatin to active ingredient is basically arbitrary and may range from 100: 1 to

1:10, especially 10: 1 to 1: 5 lie. A molar ratio in the range from 2: 1 to 1: 2 is preferred. A pharmaceutical composition according to the invention generally contains less than 10 wt .-%, preferably less than 5 wt .-%, in particular less than 1 wt .-%, unbound drug, based on the amount of active ingredient used and measured, for example, with a standard HPLC method.

The natural active ingredient may be selected from the group consisting of "camphor 1, 7, 7-trimethyl-bicyclo [2.2.1] heptan-2-one (rosemary) carvone: 1-methyl-4-isopropenyl-6-cyclohexene -2-one (caraway oil), cuminaldehyde: 4-isopropylbenzaldehyde, cumin, menthone: 2-isopropyl-5-methylcyclohexanone (peppermint oil), perillaaldehyde: (S) -4-isopropenylcyclohexene-1-carbaldehyde (cumin), cinnamaldehyde: Gammaphenylacrolein (cinnamon bark), and vanillin: 4-hydroxy-3-methoxy-benzaldehyde (spiced vanilla) ". The synthetic agent may be selected from the group consisting of "doxorubicin, salicylic acid, aminopterin, methotrexate, and nucleosides". In principle, however, any natural or synthetic active substances can be used, as long as they have reactive groups in the above sense.

The usable gelatin can basically be arbitrary, be it type A or type B, be it bovine hide, pigskin or bone. Preference is given to pigskin gelatine (type A) in pharmaceutical purity and quality.

A pharmaceutical and / or cosmetic composition according to the invention typically contains a) 0.005 to 100% by weight of at least one conjugate of a natural or synthetic active substance with a reactive group or with several reactive groups with gelatin, b) 0 to 99.99 % By weight of galenical excipients and / or excipients, c) 0 to 99.99% by weight of an active ingredient other than component a) or of several active ingredients other than component a), components a) to e) always add up to 100%.

Preference is given to component a): from 1 to 100% by weight, in particular from 10 to 80% by weight. Preference is given to component b): 0.01 to 99 wt .-%, in particular 1 to 80 wt .-%. Preference is given to component c): from 0.01 to 20% by weight.

The galenic preparation of a pharmaceutical composition according to the invention can be carried out in the usual way. Suitable solid or liquid galenic

Forms of preparation are, for example, granules, powders, dragees, tablets, (micro) capsules, suppositories, syrups, juices, suspensions, emulsions, drops or solutions for nebulization (aerosols), transdermal systems, as well as preparations with protracted release of active ingredient in their preparation conventional adjuvants such as carriers, blasting, binding, coating, swelling, lubricants or lubricants, flavoring agents, sweeteners and solubilizers, are used. As auxiliaries are magnesium carbonate, titanium dioxide, lactose, mannitol and other sugars, talcum,

Milk protein, starch, cellulose and its derivatives, animal and vegetable oils such as cod liver oil, sunflower, peanut or sesame oil, polyethylene glycols and solvents such as sterile water and mono- or polyhydric alcohols, such as ethanol or glycerol. A pharmaceutical composition according to the invention can be prepared by using at least one conjugate used according to the invention in a defined dose with a pharmaceutically acceptable suitable and physiologically acceptable carrier and optionally other suitable active ingredients, additives or excipients mixed with a defined dose and prepared for the desired dosage form.

In the context of the invention, component a) is obtainable by chemical reaction of the natural or synthetic active ingredient in an aqueous solution of gelatin. In particular, component a) may be obtainable by preparing an aqueous solution of gelatin, in particular in isotonic saline solution, in such a way that a solution of the synthetic or natural active ingredient is prepared in an alcohol, in particular in ethanol, and that the solution of synthetic or natural active ingredient introduced into the solution of gelatin and the synthetic and / or natural active ingredient is reacted with the gelatin. The mass ratio of the synthetic or natural active substance used to the gelatin used can be in the range 1: 10,000 to 1:10, preferably 1: 1,000 to 1:20.

The invention further teaches the use of a pharmaceutical composition according to the invention for the manufacture of a medicament for the treatment of a disease, which is accompanied by an altered metabolism, in particular a uptake of proteins or protein fragments to cover the cellular energy requirement of diseased cells. First, the conjugate is prepared and then the conjugate in the desired dosage form galenically prepared.

The disease can be, for example, a tumor disease or atopic dermatitis. Concrete diseases for which the The invention can be used, for example, diseases which are characterized by hyperproliferation and lack of cell differentiation, for example tumor diseases and precancerous conditions, such as intestinal tumors, gastric tumors, bladder tumors, lung tumors, or diseases, hyperproliferative

Skin disorders such as psoriasis, pituriasis subia pilasis, acne, ichthyosis, pruritus, disorders characterized by disturbance of the balance of the immune system, especially eczema.

The invention further teaches a process for preparing a pharmaceutical composition according to the invention comprising the following process steps: A) the gelatin is dissolved in an aqueous solution, in particular in a NaCl solution, B) the solution from step A) is treated with an alcoholic solution C) the gelatin and the synthetic or natural active substance are reacted with one another, D) optionally the reaction product from stage C) is mixed with galenic auxiliaries and / or excipients and / or or dissolved herein. The steps A to C are preferably carried out at a temperature in the range of 20 to 70 ° C, in particular 40 to 60 ⁰ C. In step A, NaHCO ₃ can be added.

The invention further relates (with regard to the USA) to a method for the prophylaxis and / or treatment of a disease, which is accompanied by an altered metabolism, in particular a uptake of proteins or protein fragments to cover the cellular energy requirement, of diseased cells, wherein a diseased person or a person who is about to become ill a physiologically effective dose of a pharmaceutical composition according to the invention is presented.

Finally, the invention teaches a conjugate of a natural or synthetic drug having a reactive group or having multiple reactive groups with gelatin.

The statements on various aspects of the invention always apply analogously to the other aspects of the invention.

As administration units, doses of 0.1 to 100 mg of active ingredient are possible. However, a range of 1 to 20 mg of active ingredient is preferred.

In the following, the invention will be explained in more detail with reference to examples representing only 3 embodiments.

Example 1: Preparation of a first pharmaceutical composition according to the invention

1 g of gelatin is swelled together with 50 mg of NaHCC> ₃ (about 0.6 mmol) in 20 ml of isotonic saline at 20 ⁰ C initially and brought to about 15 minutes in a water bath at 50 ° C to a clear aqueous solution.

10 mg cuminaldehyde (MW: 148.21, d = 0.981, Sigma-Aldrich, Taufkirchen, Germany, = 10.19 μl, = 67.4 μmol) are diluted with 50 μl of absolute ethanol and directly into the clear

Gelatin solution introduced. At the entry point forms a milky white haze that dissolves immediately.

The result is a colorless, clear Gelatinelosung that solidifies on cooling to room temperature.

Example 2: Preparation of a second pharmaceutical composition according to the invention

Gelatin is swelled together with 50 mg of NaHCO ₃ (about 0.6 mmol) in 20 ml of isotonic saline solution at room temperature and after about 15 min in a water bath at 50 ° C to a clear aqueous solution.

10 mg perillaaldehyde (MW: 150.22, d = 0.965, Sigma-Aldπch Taufkirchen, Germany, = 10.36 μl, = 66.6 μmol) are diluted with 50 μl of absolute ethanol and introduced directly into the clear gelatin solution. At the entry point, a milky white turbidity forms, which, however, dissolves immediately.

The result is a colorless, clear Gelatinelosung that solidifies on cooling to room temperature.

Example 3: Preparation of a third pharmaceutical composition according to the invention

1 g of gelatin is first taken together with 50 mg of NaHCO ₃ (about 0.6 mmol) in 20 ml of isotonic saline solution at room temperature swollen and after about 15 min in a water bath at 50 ⁰ C brought to a clear aqueous solution.

10 mg cinnamaldehyde (MW: 132.16, d = 1.048, Sigma-Aldrich, Taufkirchen, Germany, = 9.54 μl, = 76 μmol) are diluted with 50 μl of absolute ethanol and introduced directly into the clear gelatin solution. At the entry point, a milky white turbidity forms, which dissolves immediately.

The result is a colorless, clear gelatin solution, which solidifies on cooling to room temperature.

quality control

The conjugates obtained in Examples 1 to 3 can be tested as follows. TLC aluminum foils 5 x 10 cm, silica gel 60 F2 ₅ 4 (E. Merck, Darmstadt, Germany) are used. The eluent used is acetone, 2-propanol, water (25, 10, 5 v / v). The order volume is 2.5 μl (5% gelatin solution). The running distance is 65 mm. The chromatogram is developed with iodine vapor in an iodine chamber. The following Rf values result: all gelatin conjugates 0.00, cuminaldehyde 0.61, perillaaldehyde 0.58, and cinnamaldehyde 0.55.

Claims

claims 1. Pharmaceutical and / or cosmetic composition containing a conjugate of a natural or synthetic active substance with a reactive group or with several reactive groups with gelatin. 2. Pharmaceutical and / or cosmetic composition according to claim 1, wherein the natural active ingredient is selected from the group consisting of "camphor 1, 7,7-trimethyl-bicyclo [2.2.1] heptan-2-one (rosemary) carvone: l -Methyl-4-isopropenyl-6-cyclo-hexen-2-one (caraway oil), cuminaldehyde: 4-isopropylbenzaldehyde, (cumin), menthone: 2-isopropyl-5-methylcyclohexanone (peppermint oil), perillaaldehyde: (S) -4 Isopropenylcyclohexene-1-carbaldehyde (cumin), cinnamaldehyde: gammaphenylacrolein (cinnamon bark), and vanillin: 4-hydroxy-3-methoxybenzaldehyde (spiced vanilla) ". The pharmaceutical and / or cosmetic composition according to claim 1, wherein the synthetic agent is selected from the group consisting of "doxorubicin, salicylic acid, aminopterin, methotrexate, and nucleosides". 4. Pharmaceutical and / or cosmetic composition according to one of claims 1 to 3, wherein the gelatin is preferably porcine skin gelatin (type A). 5. Pharmaceutical and / or cosmetic composition according to one of claims 1 to 4, comprising: a) 0.005 to 100 wt .-% of at least one conjugate of a natural or synthetic active ingredient having a reactive group or having multiple reactive groups with gelatin, b) 0 to 99.99 wt .-% galenic auxiliary and / or Carriers, c) 0 to 99.99 wt .-% of a different of the component a) active ingredient or more of the component a) different active ingredients, wherein the components a) to c) always add up to 100%. 6. A pharmaceutical composition according to any one of claims 1 to 5, wherein the component a) is obtainable by chemical reaction of the natural or synthetic active ingredient in an aqueous solution of gelatin. 7. A pharmaceutical composition according to claim 6, wherein the component a) is obtainable in that an aqueous solution of gelatin, in particular in isotonic saline, is prepared that a solution of the synthetic or natural active ingredient in a Alcohol, especially in ethanol, is prepared, and that the solution of the synthetic or natural active ingredient is introduced into the solution of gelatin and the synthetic and / or natural active ingredient is reacted with the gelatin. 8. A pharmaceutical composition according to any one of claims 6 or 7, wherein the mass ratio of the synthetic or natural active ingredient used to the gelatin used in the range 1: 10,000 to 1:10, preferably 1: 1000 to 1:20, is located. 9. Use of a pharmaceutical composition according to any one of claims 1 to 8 for the manufacture of a medicament, for the treatment of a disease associated with an altered metabolism, in particular a uptake of proteins or protein fragments to cover the cellular energy requirement, diseased cells. Use according to claim 9, wherein the disease is a tumor disease or atopic dermatitis. 11. Process for the preparation of a pharmaceutical Composition according to one of Claims 1 to 8, having the following process stages: A) the gelatin is dissolved in an aqueous solution, in particular in a NaCl solution, B) the solution from step A) is mixed with an alcoholic solution, in particular a solution in ethanol, of the synthetic or natural active substance, C) the gelatin and the synthetic or natural active substance are reacted with one another, D) optionally the reaction product from step C) is mixed with galenic auxiliaries and / or carriers and / or dissolved therein. 12. The method according to claim 11, wherein the stages A to C at a temperature in the range of 20 to 70 ⁰ C, in particular from 40 to 6O ⁰ C are performed. 13. A method for the prophylaxis and / or treatment of a Disease associated with an altered metabolism, in particular a uptake of proteins or protein fragments to cover the cellular energy requirement, diseased cells, wherein a diseased person or a person who threatens to fall ill, a physiologically effective dose of a pharmaceutical composition according to any one of claims 1 is presented to 8. 14. Conjugate of a natural or synthetic drug with a reactive group or with multiple reactive groups with gelatin.