NO860012L - PREPARATIONS FOR DELAYED EFFECTS, PROCEDURES FOR THEIR PREPARATION AND SIMILAR HUMAN RESPONSE. VETERINARY MEDICINE USE. - Google Patents

Description

When using drugs on humans or animals, it is often desirable to give the actual drug a prolonged effect with suitable precautions. This problem is particularly important in the case of proteins or peptides, as these have relatively short half-lives, as they are rapidly metabolised in vivo. A typical example for this substance class is insulin, with parenteral application without the depot principle, 3-4 injections a day are necessary to prevent the diabetic from derailing.

By adding a depot carrier, however, the insulin's effect can be extended considerably.

As a depot carrier, for example, in the case of insulin, there are substance-specific delay principles, which determine the insulin's poor solubility at a physiological pH value. Additional common and general applicable forms of custody

for proteins or peptides are so-called drug delivery systems. Such systems consist of a dispensing device, e.g. a hollow body or polymer network, in which the drug is embedded and brought into the body by active or passive transport.

These systems are usually solid and are for proteins

and peptides ..for example injected or implanted under the skin. These include the methods of microencapsulation (F. Iiim, Biomedical Application of Microen-capsulation, CRCJPxess, Boca Raton (1984)), Polymerimplantate

(R- Langer and J. "Folkmann, Nature 263, 797-800 (1976), M.F.A. Goosen et al, Diabetes 32, 418-481 (1983)), erodible polymer matrices (R. W. Baker et al., Pharmaceutical Technology, 26-30 (1984)), Hydrogels (EP-A2-0092918), osmotic dispensers (US patent 3995632) or also other systems, which until now found use preferentially for low molecular weight pharmaceuticals (Selecta 21, 1966-81

(1983) , N..L. Henderson, in Topics in Chemistry and Drug

.Design, JR.C. Allen, ed (1983)).

The purpose of "drug delivery systems" is to release the drug over a longer period of time with near-zero-order kinetics. Frequently recurring problems, however, are the reproducibility of the systems, e.g. pore size and thereby the reproducibility of the characteristic, an imperfect kinetics, e.g. a kinetics of the first order with a clear peak at the beginning and a slow release towards the end of the release time, as well as compliance with activity and long-term stability of the active substance. Finally, some of the above methods are only applicable to proteins and peptides with loss of activity.

From EP-A3-0019403 it is further known to give peptides such as insulin and enkephalin a delaying effect by binding them covalently to a hydroxyethyl iron(III) complex.

From DE-A-27 58 578 and the Canadian patent 1 107 647 secretin preparations are known, which contain for stabilization glycine and smaller amounts of "Haemaccel", a gelatin partial hydrolyzate cross-linked with diisocyanates. In addition, a depot carrier of polychloroethene phosphate is described

and "Haemaccel". Furthermore, it is from US patent 4,302,448

known secretin preparations with prolonged action, such as

. contains, a .ienal.-depot.tlegeme._and in ..additional up to 10%. of a gelatin derivative^. Kragen et al. mentions in

Brit.. Med. J. 1975 III 464-466 a stabilizing effect of the addition of up to 3.5% - "Haemaccel" to very dilute insulin solutions (0.04 I.E./ml), whereby it i.a. can prevent the adsorption of insulin on the supply vessel and the tubing system in the infusion system.

EP-A-21234 mentions aqueous solutions of LHRH analogues containing 0.005% gelatin.

Aqueous insulin preparations with delayed action,

characterized vede-t content of e-t. physiologically tolerable

..- gelling agent,_has already been -suggested (German

... patent application P 3 421 613-8).

The task of the invention is to provide a simple and re-'

producible depot concept with medium-long, typically over hours to a few days persistent effect, which is generally applicable for higher molecular drugs, preferably, however, for the currently very sensitive proteins or peptides resp. their analogues, especially by their parenteral, preferably subcutaneous or intramuscular application.

This task was solved according to the invention by adding a physiologically tolerable gelling agent as a depot principle to aqueous medicinal preparations.

The invention therefore relates to prolonged-acting sterile aqueous preparations of a peptide or protein, the preparation being characterized by the fact that it contains a physiologically tolerable gelling agent in a quantity sufficient for a depot effect as the only depot principle, insulin preparations being excluded.

Gelling agents (often also called thickeners) are physiologically tolerable polymers, such as

collagen and by-products, dextrans, other polysaccharides, such as J: . e.g., levan,. gelatins.,, oxypolygelatins . ("Gelifundol"), jnodified liquid gelatins.. ("Physio-gel", gelatin partial hydrolysates., for example gelatin partial hydrolysates cross-linked with diisocyanates (polygeline, "Haemaccel"), hydroxyethyl starch or polyvinylpyrrolidone. Some of the substances mentioned are also used in colloidal plasma substitutes.

The preparations according to the invention can be liquefied or in the form of hydrogels. Surprisingly, it was found that in particular also such gel preparations which are liquid at body temperature and are therefore easily injectable, give the peptides and proteins - a delayed and possibly enhanced effect. They preferably contain more than 1%, preferably more than 2%, especially between 5 and 25% gelling agent. However, even with the addition of a smaller amount of gelling agent, form-retaining hydrogels can already form, for example the addition of approx. 0.2% agar or 0.6 gel tin is sufficient for this purpose. Above, the content of gelling agent can depend on its type amounts to 30% and more (% indications are by weight.) Especially with the smaller peptides, high gel amounts are necessary to achieve a clear depot effect.

Preferred gelling agents are gelatin partial hydrolysates crosslinked with diisocyanates (compare e.g. DBP 1 118 792 and 1 155 134), such as polygeline. They are produced

1) by reacting collagen breakdown products of a molecular weight of 2000-20000 with a smaller amount of diisocyanate than that which corresponds to the number of amino and guanidino groups present in the degraded collagen

or

2) by further breaking down the cross-linking product formed according to 1) in aqueous solution up to a molecular weight of 10,000-100,000.

The preferred cross-linked gelatin partial hydrolysates are usually added in amounts by weight of more than 1%, preferably more than 2%. In the case of smaller peptides such as sectretin, it is advantageous when the amount of this gelling agent amounts to more than 5% by weight, preferably more than 10% by weight.

The preparations according to the invention usually contain the gelling agent and the pharmacon in an aqueous medium which may additionally contain a buffer substance, a preservative, an isotonic agent and, if necessary, additional stabilizers or other common and known additives. The medium can thereby have a pH value deviating from the neutral point, however, a pH value of 3-10, especially 6-8, is preferred, because proteins and peptides are often most stable here and pH-neutral solutions are physiologically best compatible.

Unlike normal depot forms, it is therefore usually a question of clearly dissolved preparations before injection, where no homogenization is necessary. In the case of suspensions, as is known, dosing errors occur due to insufficient homogenization. However, gel-like preparations can in principle also be prepared with amorphous or crystalline active substance. Thus, it is possible to prepare poorly soluble forms of the active substance, e.g. crystals or amorphous precipitates in the form of a gel. In this way, different depot forms can be combined and ultra-long-acting "cloudy gels" can be produced.

With a sufficiently high amount of gelling agent, it is common for all preparations that under storage conditions they are solid as a hydrogel. Since the molecules can almost "be frozen" and can only slowly diffuse to the vessel wall or the liquid/air interface and then move—

..claims \and. turbulences in the gel are considerably reduced during handling, these have a special storage stability. Before use, the gels are brought, as it is

usual with ordinary insulins, at room- to body temperature, as they are liquefied, but still, in addition, compared to ordinary solutions of the active substances, the viscosity has increased.

The gel formation under the storage conditions can also be advantageous with respect to the homogeneity of the solution or the suspension. It is e.g. in the case of sedimented crystal suspensions during longer storage, it is absolutely conceivable that relatively stable crystal associates are formed which are then less easily shaken up homogeneously and can thereby lead to dosing errors. Available, however

the crystal suspension homogeneously "frozen" in a gel, then such effects can be ruled out.

As pharmaceuticals, higher molecular substances, preferably proteins and peptides for human and veterinary medicine, which are especially stable in an aqueous medium due to their predominantly hydrophilic nature, come into question. This includes, for example, hormones reps. release hormones and their analogues such as secretin, gastrin, oxytocin, differentiation factors, growth hormones, GHRH. LHRH, TRH, somatostatin, glucagon, neurotensin, clacitonin, widow-faline, ACTH, gradykinin, cholecystokinin, erythropoietin,

thymus peptides and relaxin, proteins with other functions such as immunoglobulins of different classes and types,

albumin, blood coagulation factors (eg hirudin, plasminogen activator, antithrombin III), aneurexogenic peptides, interleukins and interferons, as well as enzymes such as neuramidase, glucosidase, glycokinase, myoglobin, lysozyme and uricase.

As a carrier medium that is physiologically harmless and compatible with the active substance, a sterile aqueous one is suitable

solution, which has been made isotonic to blood in .... usual .manner, . ±. eg ... by h j elp - of ..glycerol, . table salt, glucose, and also contains one of the usual preservatives, e.g. phenol, m-cresol. or p-hydroxybenzoic acid ester in the usual amount for this purpose. The carrier medium can also contain a buffer substance, e.g. sodium citrate, sodium acetate, sodium phosphate, tris(hydroxymethyl)aminomethane. Diluted acids (particularly HC1) are used for pH adjustment, resp. lye (especially NaOH).

The physical stability of the preparations according to the invention can possibly be further increased by the addition of surface-active substances such as those e.g. is known from EP-A-..18 609, DE-A-32 40 .177, .29. 17,535 or WO-A-83/00288. The invention further relates to a method for producing a protracted sterile aqueous peptide or protein preparation, the method being characterized by adding to an aqueous preparation of a peptide or protein a physiologically tolerable gelling agent and possibly a suitable isotonicity agent, a suitable buffer, a suitable preservative and/or further auxiliaries and sterilizes the solution, for example by sterile filtration.

Likewise, it is possible to combine sterile aqueous peptide or protein preparations with sterile preparations of gelling agent and possibly further excipients.

The invention further relates to the use of the preparations according to the invention as a healing agent (for secretin preparations, e.g. for the treatment of Pancreatitis or Mucoviscidosis), the use of a physiologically tolerable gelling agent as a depot aid as well as the use of the gelling agents as a stabilizing agent when handling and storing especially hydrogels.

The active substance preparations according to the invention can be applied parenterally, especially subcutaneously or intramuscularly.... The depot effect is most pronounced with the subcutaneous method of application, but is still evident with intramuscular injection. It is therefore surprising that, alongside an extension of the effect, there is often also a strengthening of the impact. When applied intravasally, the clearly dissolved preparations according to the invention usually work quickly. They can therefore also find application in automatic dosing units such as e.g. pumps. Thereby, it is advantageous that the physiological stability is increased in many cases by the addition of gelling agents. .The invention will be explained in more detail with the help of some examples.

Eczema gel_l

Production_of_a_secre<p>olY2§liS<s>

200 g of polygeline and 2.5 g of m-cresol were at approx. 30-40°C dissolved in 800 ml of water p.i. and with IN HC1 the pH is adjusted to 3. The solution was cooled to 20°C and 100,000 KE secretin was added. HC1 with stirring. The solution was sterile-filtered through a sterile filter and filled into multiple withdrawal bottles.

Example_el_2

2 0_Y§]St_:l_dextran_60

20 g of dextran 60 and 0.9 g of benzyl alcohol were dissolved in 80 ml of water p.i., adjusted to pH 7 and 25 mg of

the LHRH analogue buserelin. The solution was sterile-filtered and filled into multiple withdrawal bottles.

Example_3

a) Production_of_a_secretion E°iY2_lIi2_:Z_: Analogous, .den;, in example. 1 - specified procedure ..it was a sterile aqueous solution with the following composition:

b) Pharmacology ske _sa^nmen^i2nin^s experiment

i) Method

Beagle dogs, for which a chronic pancreatic fistula had previously been established, and urethane-anesthetized rats, had the solution prepared according to (a) applied subcutaneously. As a comparison, a corresponding secretin preparation without polygeline was applied. The exuding Pancreatic secretion was measured in time-specific periods and the serum secretin level measured radioimmunoligally.

ii) Results

The subcutaneous application of 10 KE/kg secretol i

. the preparation without polygeline caused a 2.5 hour pancreatic secretion in rats, this effect could be extended to 12 hours by applying the peptide in the preparation with polygeline prepared according to (a) (fig. 1). Analogously, in awake dogs, the duration of action of secretol (100 KE/animal) could be extended from 3 to over 10 hours, (fig. 2). The preparation effect integrated over the duration of action was sixfold in proportion - . for submission'. of the .comparison without polyge-..line. The secretin serum kinetics correlated with the Bioassay findings.

iii) Conclusion

With the present preparation, twice daily sc. injection, a 24-hour secretory effect is maintained.

iv) Explanation_of_the_figures

In fig. 1 shows the temporal course

.of-Pancreatic secretion.on.urethane-anesthetized rats in yl/30 min. Curve 1 shows the time course of secretion after s.c. submission of lint KE/kg of

the same leveling preparation, while curve 2 shows the course after s.c. administration of 10 KE/kg of the preparation according to (a).

In fig. 2, the conditions of an awake dog after s.c. are reproduced in the same way. administration of respectively 100 KE/animal of the comparison preparation or the preparation according to (a).

Example_el_4

Prominstillin2_av<_>et<_>secretin<g>r Mucovissidosis

A depot carrier solution of the following composition was prepared: sterile filtered in the usual way and filled into sterile multi-use withdrawal bottles.

in 1 ml aliquot lyophilized in sterile glass ampoules and the ampoules fused.

Before application, the contents of a secretin ampoule were taken up in 1 ml depot carrier and then injected s.c. or i.m.

Example_5_

EE§™§£ii!i£2_3¥_§É_Sli2i§£f§ r2QE£§E§E§£_i_§£_E2iY2§i"§I carries

A solution of the following composition was prepared:

. The solution was sterile filtered over a beforehand with 0.1% polyethylene-polypropylene glycol 1800 solution and then with a placebo solution of the above composition (without interferon) pre-equilibrated sterile filter and filled in sterile multi-use withdrawal bottles.

Claims (12)

.1. Prolonged acting sterile aqueous preparation of a peptide or protein, characterized by the fact that, as the only depot principle, it contains a physiologically tolerable gel agent in an amount sufficient for a depot effect, insulin preparations being excluded. 2. Means according to claim 1, characterized in that it exists as a hydrogel. 3. Means according to claim 1, characterized by the fact that it is available in liquefied form. 4. Means according to one of claims 1-3, characterized in that the gelling agent contains gelatin, dextran, laevan, other polysaccharides, gelatin partial hydrolysates, which may be cross-linked, oxopolygelatins, hydroxyethyl starch or polyvinylpyrrolidone. 5... Means according to claim 4, .-k.--a.--r a. k t e .r 1 s e _r t in that it contains a gelatin partial hydrolyzate cross-linked with diisocyanates. 6. Means according to claim 5, characterized in that it contains more than 1% by weight, preferably more than 2% by weight, gelling agent. 7. Agent according to one of claims 1-6, characterized in that this contains a suitable isotonic agent, a suitable buffer and/or a suitable preservative. 8. Means according to one of claims 1-7, . characterized in that the active ingredient contains secretin. 9. Agent according to one of claims 1-8 for use as a healing agent. 10. Method for producing an agent according to one of claims 1-9, characterized in that a physiologically tolerable gelling agent and optionally a suitable isotonimide agent, a suitable buffer and/or a suitable preservative are added to an aqueous preparation of a peptide or protein and the preparation is sterilized or a peptide or protein is added to an aqueous preparation of physiologically tolerable gelling agent, which possibly contains a suitable isotonic agent, a suitable buffer and/or a suitable preservative and the preparation is sterilised. 11. Use of a physiologically tolerable gelling agent as depot aid in aqueous preparations of peptides or proteins, excluding insulin preparations. 12. Use of a gelling agent as a stabilizing agent - wood. storage - of . hydrogels according to requirements .2.