NL1002400C2 - Oral calcitonin preparation and method of recovering calcitonin. - Google Patents

Description

Title: Oral calcitonin preparation and method of recovering calcitonin

The invention relates to an oral calcitonin preparation, as well as a method of obtaining calcitonin and products containing calcitonin.

5 Calcitonin is a polypeptide hormone containing 32 amino acids. It is produced in mammals in the parafollicular cells of the thyroid gland (thyreocalcitonin); and in birds and fish in the ultimobranchial body.

Calcitonin is used therapeutically, for example in the treatment of Paget's disease, in hypercalcaemia and (post-menopausal) osteoporosis. It inhibits the action of osteoclasts and promotes the clearance of calcium and phosphate in the urine. In addition, calcitonin has been reported to reduce pain symptoms associated with these conditions.

Because of the therapeutic application, there is a great need for calcitonin. A drawback, however, is that it can only be obtained through complicated and / or high cost-involved processes.

Calcitonin as a natural product is mainly obtained in pure and very active form from the ultimobranchial glands of salmon, eel or poultry.

According to European patent application 0 606 816, calcitonin is obtained from salmon by specifically binding it to columns and then purifying it. The calcitonin thus recovered is 40 times more active than calcitonin of human origin.

EP-A-0 146 842 discloses that calcitonin 30 is recovered from the ultimobranchial poultry glands by acid extraction followed by a number of purification steps using hydrophilic organic solvents, cation exchangers and gel chromatography. The activity of this recovered product 10024002 was found to be equal to or stronger in action than calcitonin recovered from salmon or eel.

Synthetic analogs and products obtained by recombinant DNA techniques have also been described.

Calcitonin is used in both natural and modified form.

For example, European patent application 0 347 105 describes that cysteine sites in the polypeptide are sulphonated, while according to international patent application 95/18152 combinations with (poly) cyclic compounds should be pursued.

European patent application 0 261 552 describes the production of calcitonin via recombinant DNA techniques.

As is well known, the simplest and least invasive pathway for patients to administer a pharmaceutically active agent through the gastrointestinal tract is to take the pharmaceutical orally. However, this application pathway is not always equally efficient for polypeptides such as calcitonin. Intestinal enzymes, mainly proteases, degrade the polypeptide to smaller peptides and amino acids, which degradation products are inactive.

For this reason, the known derivatized or modified or modified calcitonin preparations have hitherto been mainly administered orally by inhalation, injection or vaginal or rectal application.

The fact that in order to achieve a sufficient effect, calcitonin must be administered regularly, the latter methods of application have many drawbacks. The most suitable application route for patients is the oral route.

Proposals have been made to improve the recording efficiency of oral preparations.

Thus, Japanese publication 93-274737 describes special calcitonin derivatives with an improved enteral absorption. For this purpose, fatty acid groups or lower alkoxy carbonyl groups are linked to the polypeptide chain. The 1002400 3 derivatives are suitable for the treatment of pain due to osteoporosis and hypercalcaemia, and improve the onset of bone mass.

European patent application 0 535 827 proposes the incorporation of calcitonin in fat-like (emulsifier) material and the encapsulation of calcitonin in gelatin. These calcitonin products are better absorbed without the need for an absorption enhancer.

Furthermore, Japanese publication 62-010020 describes a composition containing, in addition to calcitonin, a fatty acid ester with an HLB of 11-16. This composition is said to have improved absorption behavior.

International patent application 87/00750 further describes for the same purpose a method in which calcitonin is surrounded with a thin phospholipid skin, thereby bringing it into a liposome form.

The proposals for orally suitable calcitonin formulations all require relatively expensive modification steps to necessarily protect calcitonin from rapid inactivation in the gastrointestinal tract. In particular, pure calcitonin is always used as the basic raw material, which has been obtained via complicated and / or high-cost-entailing technologies.

It has now been found that calcitonin is present in egg shells and can be extracted therefrom. In addition, eggshell calcitonin can be efficiently administered orally without the need for an additional derivatization or other modification step.

The invention therefore relates in a first aspect to calcitonin derived from eggshells.

In addition, the invention relates to a method of obtaining an orally applicable calcitonin product, wherein calcitonin is released from egg shells. For this purpose, for example, calcitonin can be incorporated in an aqueous medium or an aqueous solvent system, but also in other solvents 1002400 4 in which calcitonin dissolves in contrast to the inorganic eggshell matrix. In addition, calcitonin can be obtained by at least partially dissolving the eggshell matrix.

5 Egg shells, with or without membranes, form an ample stock of waste product. In order to optimize the extraction of calcitonin, the eggshells are first ground, preferably into the finest possible eggshell flour. It is not necessary to remove the 10 fleece remnants from the eggshells.

The egg shells can optionally be dried before being ground, which facilitates the grinding operation. In addition, the starting product can be pre-sterilized without the activity of the calcitonin decreasing unacceptably. Apparently, the calcitonin is sufficiently protected by the lime matrix of the eggshell.

A very great advantage of the method according to the invention is that no extensive purification steps of the calcitonin product are necessary. It is not a problem, and even an advantage, if not all residues of the eggshell matrix are removed. By administering calcitonin in combination with a lime product, the effect of calcitonin is greatly enhanced.

It is believed that one of the reasons why calcitonin from eggshells can be used orally lies in the fact that this calcitonin form is protected from degradation under the influence of enzymes or juices by complexing with components of eggshells or by embedding in the eggshell matrix. in the gastrointestinal tract.

In another aspect of the invention, calcitonin can be administered by orally administering eggshells in any form. Eggshells contain calcitonin in a form and amount that the digestion in the gastrointestinal tract can be sufficiently avoided or inhibited, and that the calcitonin can be incorporated into the system and have the desired effect there.

Experiments (vide infra) have shown that the calcitonin products according to the invention give a physiological pain-reducing effect when administered orally. In addition, these products can be used in all mammals suffering from a calcitonin treatable condition. Furthermore, the calcitonin product of the invention can be used prophylactically.

The invention further relates to an oral preparation containing calcitonin obtained from eggshells.

Incidentally, it is known from the international PCT / NL / 95/00396 of the present group of inventors, that milk products enriched with eggshell flour give rise to an improved calcium absorption. Moreover, it is described herein that additional advantages are obtained with eggshell flour because of the presence thereof. magnesium compounds. It is also pointed out that it may be useful to add vitamin D sources.

In addition, Arch. Ger. Geriatr. (1991) 487-490, it is known that administering eggshells can lead to a pain-relieving effect.

However, neither publication describes or suggests that in fact calcitonin is responsible for the effects.

Preferably, a magnesium source is also present in the oral preparations according to the invention. This magnesium contributes to the pain-reducing effect of the calcitonin. In a preferred embodiment, the calcium to magnesium ratio in the composition of the invention is between 3: 1 and 12: 1. Both the calcium and the magnesium can come from eggshells and / or from external sources. This ratio is so selected for physiological reasons, namely to effect optimal calcium retention in bones.

1 o o Z A 0 0 6

It is noted that in Magnesium Res. 2. (1989) 142, it has been described that magnesium, when administered to osteoporosis patients, has a pain-relieving effect.

Further details about the amounts of calcium and magnesium are given in the aforementioned international patent application PCT / NL / 95/00396.

It is furthermore advantageous to combine the preparations according to the invention with components which are known to positively influence the calcium retention, in particular with vitamin D or by combining the preparation with a source rich in vitamin D. Other examples are the vitamins A, K and / or C, sources of fluoride, vanadium, boron and / or silicon ions, glycine, and casein phosphopeptides.

The oral preparation according to the invention can be used in any known orally applicable form. Preferably, the preparation is in the form of a pill, a swallowing or chewing tablet, or a liquid preparation. In the latter case, by selecting a suitable liquid medium in the method according to the invention, the directly obtained product can be administered.

In addition, the calcitonin obtained from eggshells can be included in a food or used as a dietary supplement, for example, for prophylactic reasons.

The invention further relates to a method of administering calcitonin, wherein an eggshell product is administered orally.

Finally, the invention relates to the use of eggshells as a source of calcitonin.

The invention is now illustrated by the following non-limiting examples.

Example 1 Finely ground eggshell flour obtained as described in international patent application PCT / NL / 95/00396 contained about 38 wt.% Calcium, 0.4 wt.% Magnesium, 0.1 wt.% Phosphorus and 0.6 wt. .% nitrogen.

1002400 7

This product was suspended in 90 ml water in an amount of 10% and tested for calcitonin. In the suspension, the calcitonin content was determined according to the standard method of Nicholson (Nicholson Institute).

The suspension contained 0.09 μg of calcitonin.

Example 2

Capsules were prepared for oral use containing 0.7 grams of fine calcitonin containing eggshell flour and 0.05 grams of a mixture containing sodium ascorbate, zinc lactate, vitamins E, D3, B6, K, B2, B1, B12, silicon dioxide, boric acid and milk protein ( casein, whey protein).

15 Example 3

The product was prepared according to Example II, but 0.7 grams of calcitonin-containing eggshell flour was replaced by 0.5 grams of calcitonin-containing eggshell flour and 0.2 grams of magnesium hydroxycarbonate.

20

Example 4

A powdered product was prepared by mixing reduced-fat milk powder with a mixture of eggshell flour and maltodextrin, dried together on a wheel dryer, as well as a vitamin-mineral premix. The product contained per 100 grams 21 g of milk protein, 14 g of fat, 2.2 g of calcium, of which 1.5 grams came from fine eggshell flour and 0.6 grams of magnesium. Via the vitamin-mineral premix, as micro-30 ingredients, iron, zinc, copper, iodide, vitamins A, C, D, E, K, BI, B2, B6, B12, folic acid, niacin, pantothenic acid, biotin were added.

The Ca / Mg ratio was <4: 1.

35 Example 5

The product of Example 4 but the reduced-fat milk powder was replaced by a mixture of cereal flour and soy.

100 * ^ 00 8

Example 6

The product of Example 4 was given for 4 months to subjects with proven osteoporosis, who had pain complaints, which had hitherto been treated with conventional pain killers.

None of the participants used hormone preparations (eg estrogens), bisphosphonates, or other preparations that could be thought to influence pain.

Per day, the subjects took in 3 g of calcitonin-containing fine eggshell flour with the product.

The pain perception was followed by specialists who followed the pain in the back or bones, the increase in pain after 15 exertion, falling asleep due to pain on a classification scale of 1-4 (1 = no complaints; 2 = a few complaints; 3 = quite a few complaints; 4 = very bad complaints).

In addition, the extent to which the use of 20 painkillers was discontinued or continued was monitored.

It was concluded that after 4 months the product had significantly reduced pain symptoms in 73% of cases.

A similar result was achieved with the administration of 6 grams of calcitonin-containing eggshell meal to test subjects alone.

100/400

Claims (9)

1. Calcitonin available from eggshells. 2. A method of obtaining an orally applicable calcitonin product, wherein calcitonin is released from egg shells. 3. Oral preparation containing calcitonin obtained from egg shells. The preparation according to claim 3, which is enriched with a magnesium source. 5. A composition according to claim 3 or 4 enriched with a vitamin D source. A preparation according to claim 3, 4 or 5, in the form of a pill, a swallowing or chewing tablet, or a liquid preparation. 7. Preparation according to claim 3, 4 or 5, included in a foodstuff. A method of administering calcitonin, wherein an eggshell product is administered orally. 9. Use of egg shells as a source of calcitonin. 100 * 400 ΠΛΓΓνπ I DC I ncrrcnuc NEWS1STUDY OF INTERNATIONAL TYPE OF IDENTIFICATION OF THE NATIONAL APPLICATION Kenmei * of the applicant or of the gemnogoe _ NW 9056 NtMiuneu application no. Filing 1002400. February 20, 1996 Claimed priority applicant (Name) FRIESLAND BRANDS B.V. Reconcile a tier for oen onoerzoe * of mumaaanaa type Ooor do insanee for Inamatonaai OnurzoeK (ISA) to reconcile for a research of «emaeonaiai type acc. No. specify) according to oe mamaoonaie cassalicaae (IPC) Int. Cl. 6: C 07 K 14/585, A 61 K 38/23, A 61 K 47/04 II. FIELDS OF TECHNIQUE RESEARCHED Untouchable mmtmum documentation__j Classification system Classifieanesvmimi _ | Int. Cl.6 C 07 K, A 61 K Unsurpassed aocurnenaoa other than the minimum documenaoe insofar as very detailed documents are included in the research »prayers sentence * ^ III. i ~ NO RESEARCH MAY BE PERFORMED FOR CERTAIN CONCLUSIONS (comments on supplement solad) ivi fV.! LACK OF UNITY OF INVENTION (oomerwngen on replenishmentolad) j J -: - 'om PC7.1SA / 201laiC £ t9Si