DE1617780B2 - Scarring agents - Google Patents

Description

The invention relates to a masking agent that preferably suitable for topical applications on the skin or mucous membranes

Collagen fibers have already been used to manufacture used for surgical sutures and ligatures and for the manufacture of replacement parts for blood vessels.

From US-PS 31 57 524 means based on collagen are known, in which, however, the collagen through Treating with a strong base and dehydrating at elevated temperature is in denatured form The denaturation of collagen means the destruction of certain functional groups in the Molecular structure of collagen protein. The use of tanning agents described there, such as formaldehyde, leads to denaturation. This is shown by the later-described comparison between one after Sample B obtained in this prior art with a sample A according to the invention.

The same also applies to the collagen foam described in FR-PS 14 41817, which consists of Non-fibrous bone glue is made by swelling with acid, with the pH increasing The addition of NaOH is adjusted to 7. The addition of this strong base in turn makes the collagen denatured The product obtained after lyophilization is further because it is completely dehydrated denatured and no longer shows any moisture content. Due to the present in sponge form Collagen only has a hemostatic and mechanical protective effect. - This also applies to the product described in FR-PS 14 86 237, the is also a product of degradation or denaturation of collagen after it has been exposed to Has been subjected to tanning as the later-described comparison between Samples C and Sample A shows.

The type of collagen described in Virchows Arch. Path. Anat. 339, 198-205 (1965) is in this Publication not identified, but it can be seen from the statement that the use of heterologous collagen had no favorable result on the development of injuries, conclude that in this case too denatured collagen was used. Such collagen will not have a scarring effect mentioned

It is therefore the object of the present invention to provide an agent with scarring properties and good To create tissue compatibility.

According to the invention it has now been found that certain ίο KoUagenformen, namely gently insulated collagens, represent extremely effective scarring agents with good tissue tolerance.

The subject of the invention is thus a marring agent, obtainable by

a) fibrous, made from the hide of cattle, horned animals or Adds water to unpaired, not noticeably altered collagen produced, b) this mass is shaken in a dilute acidic medium until an aqueous gel is formed and

c) brings this gel into the desired pharmaceutical form, with the hair coloring agent if it is in the form of a sheet or plate, has a water content between 5 and 10% and optionally contains a plasticizer

According to a preferred embodiment, the marring agent according to the invention is characterized by this from being in the form of a compact, fibrous collagen sheet or plate, or in the form of a non-compact collagen sheet of airy structure is present, e.g. B. in the form of a gauze, compress or one Association.

The agent according to the invention can therefore be in various forms, both for the local Application as well as for oral, intramuscular or intravenous administration are suitable. So it can one the form of ointment, cream, powder, foam, eye drops, gynecological capsules or tablets, Have suppositories, tablets, solutions, etc.

Self-adhesive bandages can also be produced

are made by combining a collagen and

z. B. an adhesive strip, which the

Holds collagen in place Furthermore, the marking agent contains

men with the collagen at least one other pharmacodynamically active principle. Except for his role of a scarring agent, the collagen then plays the role of a carrier for this other active principle. That Collagen can be the only vehicle used. According to a further embodiment, however, a other carriers can be combined with the above system.

The said active principle is an antiseptic agent which z. B. selected from the following substances

j5 can be: eosin, crystal violet, hexachlorophene,

Azulene and Gentian Violet A combination with one Anliseptic is natural to topical application

of the scarring agent

The pharmacodynamically effective principle can be like

the collagen be a scarring agent, e.g. B. coal tar, or an agent for the treatment of skin diseases, which e.g. B. cade oil or ichthyol can be.

In general, the pharmacodynamically active principle can be any one known per se. Be it

h5 noticed that the union of collagen and

Cortisone derivatives, e.g. B. fluorocortisone, especially

is of interest for the treatment of certain dermatoses.

The fibrous sheet or sheet which the

Can contain pbarmakodynarnisch active principle or not, may contain an agent which corresponds to the The sheet or the plate is given a suppleness This agent is preferably glycerine, which allows the collagen to be made pliable, since the Preparation of the same generally results in the removal of all accompanying solids

The scarring agent of the invention has at Its topical application on the skin has a moisture content between 5 and 10%. This salary corresponds to the maximum swelling capacity of the collagen fibers.

The sheet or plate of fibrous collagen is preferably prepared by printing out an aqueous gel from fibrous defatted collagen and then obtained by drying the squeezed out layer.

Below is a method of making the scarring agent of the present invention and an example of use the same described for a healing treatment.

Manufacturing

A collagen that is not noticeably changed is used, which is obtained from the skin or skin waste of cattle, This collagen is fibrous and can contain water absorb so that it gradually turns into an aqueous gel

This fibrous collagen is e.g. B. previously dried Calf collagen, which has the following properties:

- Humidity:
between 5 and 10%

- related to the completely dry product Total nitrogen content:

15 to 17%

- nitrogen of the amide functions:
0.5%

- sulphurous ash:
5 to 6%

'This collagen can contain various amounts of salts, such as sodium chloride, ammonium sulfate etc., which remain in the final product when following the manufacturing process below is proceeded. 300 g of this fibrous calf collagen (with the above numbers appropriate moisture content, i.e. between 5 and 10%) and 101 water, if necessary with an addition of acetic acid (1 to 3%). This addition of acid causes a better effect in certain cases Swelling the fibers. It should be noted that this fibrous collagen is defatted so that it can be used for production A plasticizer may be added to certain forms of the final scarring agent, e.g. B. glycerin. In this case z. B. 100 g of glycerin added to the jar.

It is shaken in vacuo until a gel is obtained, which takes about 6 hours. Then that will Let the preparation stand for about 12 hours.

The aqueous gel thus obtained is then pressed out through a nozzle, the opening of which can be in the form of a slit of variable width. This gives a layer which ^{is dried in an oven at 50 °} C. in a vacuum. This layer turns into a sheet or plate, which the glycerine gives a great suppleness. The drying continues until the remaining water content is between 5 and 10%.

The incorporation of the glycerine also has the consequence that the absorption of the wound fluid (which the scarring) is prevented, in contrast to e.g. wadding,

The thickness of such a sheet or such a plate is, for example, between Via mm and 2 mm.

If other pharmaceutical forms should be produced, ζ, Β, ointments or creams, the Treatment at the stage of aqueous gel ai.f that ίο then using the usual techniques in the desired pharmaceutical form is brought.

If another pharmacodynamically active principle

to be combined with the collagen of the sheet or plate, the above preparation is made modified as follows: before shaking the active principle in the mixture of water,

Collagen and optionally glycerin dissolved or

suspended If necessary, a dispersant is

z. B. polyethylene glycol, added to homogenization to make the whole thing easier.

The amount of scarifying agent will of course depend on the intended application. At a dermatological use must be kept to the concentrations that are usual for this various active principles (e.g. antiseptics) can be used.

Application example With the scarring agent according to the invention

jo was treated a patient with a leg ulcer. This ulcer had a diameter of three inches and sat on the front inner surface of the right leg about 8 inches from the top of the Ankle removed This ulcer occurred in a woman who had varicose veins on her legs and had phlebitis after conceiving at the age of 23. The ulcer had for the first time about two years before starting to use the scarring agent according to the invention educated

After an outpatient treatment (Unna boots) the ulcer had healed relatively quickly during the start of treatment About 1 cm in diameter. Six months after the first treatment, the ulcer came back and had since gradually had periods of incomplete healing during either outpatient treatments or, on the contrary, continue with the local application of various ointments during rest Spread over this ulcer, which had been developing for more than a year, were plates of Collagen with eosin (23% eosin) and then plates of Brought collagen without eosin. Complete scarring was achieved after three weeks.

From the beginning of the application, the formation of fleshy outgrowths was observed, what the Healing process testified.

This plate had a thickness of about ½ to mm. These various findings show the very large

m> effectiveness of the scarring agent according to the invention as well as its complete compatibility with the skin and the underlying tissues.

The following are the test results based on the X-ray analysis / ur structure of the collagen

<> 5 form according to the state of the art (based on Examples 1 of US Pat. No. 3,157,524) and the collagen form of the scarring agent according to the invention are described.

I, chosen procedure

The procedure is described in detail in the following publications:

1, "Collagene" Ramanathan N "Inter Science Publishers, New York and London, 1960 (p. 117," X-Ray diffraction pattern of collagen «).

2. "Treaty on collagen", Volume 1, Chemistry of collagen. Ramachandran G. N. Academic Press, London and New York, 1967 (p. 110 "X-Ray diffraction pattern of collagen and its interpretation").

The degree of denaturation or degree of degradation of the natural collagen molecule can be determined by means of an X-ray diffraction study using a comparison sample (in this case the collagen sample A according to the invention). In the case of a series of samples to be compared with one another, an analysis was carried out on 100 mg samples which were compressed ^{at 6 kg / cm 2;} Counting: 1 meter - every 0.1 ° - use of a rotatable membrane sample carrier (speed: 4 rpm) - horizontal goniometer.

The examination of the collagen fiber structure by means of X-rays is mainly based on the determination of the areas of the peaks P \ and Pi (especially Pi) of the curves, which show the number of strokes per minute depending on the value of the diffraction angle. The peak P \, which corresponds to a diffraction angle of approximately 74 °, gives the diameter of the triple helix of the collagen molecule. This diameter is generally between 12 and 16 Å, depending on the degree of hydration of the collagen molecule. The peak P ₂ corresponds to a diffraction angle of approximately 312 "and represents the distance between the amino acid residues inside a chain and sagarically between two chains, ie approx. 235 A.

For a given series of measurements, the area of each peak P 1 or P ₂ (in particular Pi) is a growth function of the number of non-denatured molecules (ie the number of molecules with a helix structure). In this way, the degree of denaturation can be determined with the aid of the area.

The table below relates to a series of measurements carried out with two samples A and B; the areas of the peaks P \ and Pi of the above-mentioned respective Versuchskurveti were determined planimetrically

The collagen A used as a reference sample consists of collagen according to the invention.

The Kotlagen B is such according to the US-PS 31 57 524.

II. Results obtained

sample

Area
by P \

Area
from Pi

259.8
193.5

40.1
16.3

IH. Obtaining the collagen B

This collagen was obtained as follows:
Defatted and carefully cleaned veal tendon

s was divided into tiny leaflets, which then become one were subjected to enzymatic treatment (0.15 parts of ficine and 3.75 parts of tetrasodium ethylenediaminetetraacetate In this amount of enzyme solution, 75 parts of tendon were placed for one night. The remainder

ίο Ficin was made by adding 2 pounds to 5 parts of a 30% hydrogen peroxide solution destroyed.

To this mixture of tendon lamellae in approx. 750 parts of water, 2,244 parts of water were added and 537 parts of cyanoacetic acid were added.

The solution was cooled to ° under 25 C and then centrifuged at about 60 rev / min, 3 hours after homogenizing and filtering the collagen dispersion was poured into stainless steel molds and in a night at - 20 ⁰ C cooled Then, the gelled blocks were removed from the molds and placed in a room temperature circulating bath containing 121 99% isopropanol and 25 ml of concentrated ammonium hydroxide

comment

This table shows that the B collagen is about 60% denatured.

Experience shows that "collagen" B is not collagen. but by breaking down the collagen molecule arises. Such a product should be referred to as a "collagen-like product".

This preparation is generally prepared as in Example 1 of US Pat. No. 3,157,524, but for comparison purposes using the raw material which is also used for the production of the collagen A according to the invention is used.

Comparative therapeutic experiments with the inventive method are described below Collagen A and the well-known "collagen" B described above were carried out.

These collagens became smaller for treatment Varicose ulcers used after cleansing and an antiseptic treatment in the event of one Any bacterial proliferation, these ulcers no longer developed and did not show any Scarring tendency. Any important arterial or venous factors that would have falsified the results can have been turned off. All patients were treated under the same conditions, especially with regard to rest. The collagen was localized to the area of the ulcer upset.

I. Therapeutic trials using
an ointment (with topical, daily use)

The ointment had the following composition:

- Collagen A or Collagen B:

Three parts by weight (calculated on dry matter)
5j - glycerine:

10 parts by weight

- Vaseline:

1 part by weight

- polyglycol stearate:
3 parts by weight

- demineralized water:
94 weight parts.

1. Result obtained with collagen A.

After one day of treatment: The surface of the The wound was full of fleshy growths and the edge of the skin was healthy. After a 2 month treatment period the scarring was practically complete.

2. Result obtained with collagen Ii

After 2 months of treatment, the ulcer looked practically unchanged. No fleshy outgrowths whatsoever. Good compatibility.

II. Therapeutic attempts using

a plate (when used locally,

3 χ renewed weekly)

The collagen plate A was obtained according to the invention. The collagen plate B was obtained by molding the collagen B, the preparation thereof above is described.

1. Result obtained with collagen A.

15 days after the start of the treatment, the fleshy growths closed the wound more and more, and Skin began to form. After 7 weeks it was

1 ICIIUII5 | / l ClfMIJ \ .ll ai / gbJUllUMVll.

2. Result obtained with collagen B.

After 7 weeks of treatment, some fleshy outgrowths appeared, the edges of the ulcer however remained destroyed and no skin formation took place.

III. Therapeutic trials using
of a powder (with daily topical application)

The powder used in the experiments was made by mechanically crushing the plate according to previous attempts.

1. Result obtained with collagen A.

After 15 days: Excellent granulation of the Connective tissue, then skin formed around the edges of the wound. After 2 months it was Scarring practically complete.

2. Result obtained with collagen B.

After 2 months of treatment, the ulcer remained slack and showed no tendency to flesh; the diaper remained clean but had a large area of inflammation around the ulcer.

To compare the scarring agent according to the invention with the product of FR-PS 14 86 237 was this is made as follows:

A collagen gel was made from pig's house parts that were treated with lithium hydroxide, as in the example (A and B, page 4) of FR-PS 14 86 237 described. The gel thus obtained was in the form of a sheet

ι brought and with a 4% glutaraldehyde solution tanned, then treated with acetone, whereupon the sheet obtained is placed on a cover sheet, prepared as below C of the example given in the citation.

1 »The test described above was carried out with this product (sample C); the results (expressed as a comparison with sample A from collagen according to the invention; see table above) are the following:

sample

Area of P \ Area of P ₂

250.8

40.1

The table above shows that the collagen of sample C is more than 70% denatured or degraded and therefore no longer represents collagen at all. This denatured product actually exists to

> s large part of gelatin.

The topical application of the product of Sample C to a patient with a small ulcer am right B - ': r of the kind according to the invention with collagen according to the experiment cited above was treated, has not produced any positive net result after two months of treatment (missing Epithelialization); the dressing made with sample C was renewed every week (patient, age 40 years). For two more months each was

j? Treatment interrupted; the ulcer had closed practically undeveloped; then a collagen ointment according to the invention was applied (an application every week for two months); after four weeks, flesh growths appeared as a sign of good Epithelialization process and the ulcer was practically healed by the end of this treatment.

The treatment was preceded by an antiseptic treatment to avoid any bacterial growth to stop and / or prevent; the patient showed

· »> No arterial or venous factor affecting the Could have falsified results.

Claims (2)

Patent claims: 1. Vemarbmittel, obtainable by the fact that a) fibrous, made from the hide of cattle, horned animals or adds water to collagen that has not been noticeably changed, b) this mass is shaken in a dilute acidic medium until an aqueous gel is formed and c) brings this gel into the desired pharmaceutical form, wherein the pharmaceutical agent, if it is in the form of a sheet or plate, a water content between 5 and 10% and optionally contains a plasticizer 2. Vemarbmittel according to claim 1, characterized characterized in that it is in the form of a compact, fibrous collagen sheet or plate, or in the form of a non-compact collagen sheet of airy structure.