DD156082A5 - PROCESS FOR THE PREPARATION OF ACTIVE CHONDROITIN SULFATE - Google Patents

Abstract

The invention relates to a process for the preparation of active chondroitin sulfate A and C or a mixture thereof. The aim of the invention is to provide an improved process for the production of biologically and physiologically "active" chondroitin sulfate in pharmaceutical grade and good yield. According to the invention, an aqueous solution is prepared from a starting material, for example bovine tracheal cartilage or fish tissue. To this solution is added a complexing agent, for example a quaternary ammonium salt, to form a precipitate, which is a water-insoluble complex of the active material and complexing agent, and the complex is split into pharmaceutical grade to recover the "active" material.

Description

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A process for the production of _r ^ aktivern chondroitin sulfate

The invention relates to a process for the preparation of active chondroitin sulfate A or active chondroitin sulfate C or a mixture thereof

The compounds according to the invention are used as medicaments, for example, for the excitation of collateral circulation in the forth rich _s supplied through the branches of the coronary arteries v / ground of limitation of cardiac conditions such as myocardial infarction, acute coronary insufficiency with acute myocardial ischemia "

The present invention is a partial further treatment of the patent application with the application filing no. 82 * 045 filed on 5 October 1979.

Characteristic of the known technical solutions

As in US no. 3 "895" 106 and the US-PS Ur. No. 3,895,107, there is a method of treatment for inhibiting the development of atherosclerotic diseases in animals belonging to the mammalian species, in which the collateral circulation is stimulated in areas of the heart supplied by the branches of the coronary arteries and the occurrence of cardiac conditions such as myocardial infarction, acute coronary insufficiency with acute myocardial ischemia in humans with coronary heart disease consisting in the substantially regular and prolonged oral administration of an effective amount of biologically and physiologically "active" chondoitin sulfate A, "active" chondroitin sulfate C or mixtures thereof to mammals the activity is manifested by the fact that the

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Plasma thrombus formation time is extended by at least 80 % in rabbits 6 to 12 hours after administration, as described in the Chandler loops ~ method *

From a recent study comparing oral chondroitin sulfate with the placebo effect in preventing postoperative deep vein thrombosis, it was concluded that oral CSA reduces the incidence of postoperative DVT, such as by the radioactive fibrinogen assay in general surgical patients In addition, the investigation has shown that no complications occurred by administering the drug

By known methods, the "active" CSA and CSC are recovered by digesting and solubilizing the ground and defatted starting material, such as bovine and shark cartilage, by prolonged exposure to papain activated by cysteine hydrochloride and disodium versenate. "To the solution, the various foreign material together with the required "active" CSA and / or CSC, two volumes of acetone are then added, after which the desired CSA and / or CSC is precipitated. "

This described procedure has some disadvantages in practice. Precipitation is not complete unless large amounts of acetone are used. "The use of large quantities of acetone is inconvenient, hazardous, and expensive, and even at best, at least some of the desired active CSA and / or CSC

«. *,, -, -. ϋ 21.7-1981

58 661 18

not precipitated and lost · The more the relative volume of the acetone is reduced for easier handling, the greater the loss.

The aim of the invention is to provide an improved process for the production of biologically and physiologically active chondroitin sulfate in pharmaceutical grade and good yield.

Presentation of the .Wesens of the invention

The object of the invention is to find suitable complexing agents for the separation of chondroitin sulfate.

According to the invention, complex-forming compounds such as quaternary ammonium compounds selectively form water-insoluble complexes with "active" CSA and CSC. They cause substantially quantitative precipitation of the CSA and CSC, with all other undesired substances remaining in the solution at the same time.

With the complexing agents according to the invention, the difficulties and problems resulting from the use of acetone are eliminated.

According to the invention, it is no longer necessary; Moreover, it is quite surprising that these complexes form and that they selectively form with the "active" CSA and / or CSC. Although not supported by theory, it is believed that the complexing agents and

21.7.1981 58 661 18

the "active" CSA and / or CSC are oppositely charged and therefore attract each other so that they form such a type of binding. In any case, the effect of the bond is that a complete precipitation is achieved. The binding can then be readily cleaved by a base such as sodium hydroxide and / or hypertonic saline to obtain the free "active" CSA and / or CSC. It is believed that this improved method represents an important advance in the production of this important drug.

The method according to the invention can be used for "active" CSA and / or CSC from all conventional sources, such as bovine porcine, whale and shark trachea, swine rachea and of course mammalian tissue in general.

The supply of animal or fish raw material, such as that supplied by warehouses, involves the cutting off of fat and the cutting or grinding of the trachea or other tissue.

The finely divided starting material is then digested using pepsin, papain or other proteolytic enzymes to remove the protein. Simultaneously or sequentially, the protein removal and solubilization of the CSA and CSC can be carried out with the aid of a strong base such as sodium hydroxide and potassium hydroxide. Strong hydrochloric acid is also well suited for this purpose.

In any case, an aqueous solution of CSA and / or CSC is obtained. This solution contains many other undesirable, and in some cases detrimental, components derived from the chemical digestion of the starting material, and it

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It is a principal object of the invention to separate and remove "active" CSA and / or CSC from these undesirable or harmful substances. "

According to the invention, a novel and unique insoluble complex of CSA and / or CSC and complexing agents is formed from aqueous solutions. The complexing agents used are, for example, cetylpyridinium chloride or n-alkyldimethylbenzylammonium chloride having the formula:

CH.

CH,

wherein the alkyl group is predominantly

-Q

is. Such

Substances are available under the name "Marqual".

Another and equally useful complexing agent for "active" CSA and / or CSC are various anion exchange resins such as "Dowex 1-X". These fabrics are described in more detail in U.S. Patent No. 2,591,573 and U.S. Patent No. 2,591,574. In general, these resins are crosslinked benzene-divinylbenzene copolymers containing lower alkyl quaternary amine groups.

After precipitation and collection of the complex, it may be disrupted or disrupted by mixing it with a mild base and / or hypertonic saline solution in conformation.

^w 21.7.1981

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This material can be used directly in a capsule, tablet, etc. for oral administration to humans, especially as antithrombotic agents. Dosage levels are as described in the above patents. If the drug is to be administered in parenteral form, the released "active" CSA and CSC may be further treated with oxidizing agents such as potassium permanganate to eliminate any traces of harmful oxidizable components.

Ausführungebeispiel

The following example is illustrative and not intended to be limiting in any way. In the examples, parts and percentages are given by weight unless otherwise stated.

4.5 kg (100 pounds) of prepared bovine rachea was cut into 25 mm (1 inch) pieces and added to approximately 190 liters (50 gallons) of deionized water in a container. The pH was increased by the addition of approximately 400 ml of glacial acetic acid about 4 »5 set. The resulting suspension was stirred, raising the temperature of the contents of the container to about 50 ^° C. 567 g (1 .1 / 4 pounds) of pepsin were added and stirring was continued for 30 minutes. In the container a further 190 1 (50 Galionen) were passed deionized water ', and mild stirring was continued for 12 hours at 50 ⁰ C until the trachea cartilage was freed from connective tissue. The temperature of the suspension was then increased to 80 ⁰ C, and on

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The top of the fluid formed a layer of fat. The digester liquid was withdrawn through a drum centrifuge and discarded *

The remaining Peststoffe were washed twice with 190 1 (50 gallons) of hot water (60 to 80 ⁰ C). A sodium hydroxide solution was prepared by adding 680 g (1.5 lbs) of NaOH to 19 l (5 gallons) of deionized water in a container. The twice-washed pesticides were placed in the sodium hydroxide solution and the volume was adjusted to 45 liters (12 gallons) with deionized water and the pH to 9-10. The container contents were stirred for 12 hours at 37 ^° C. The value was then adjusted to 6.5 with the aid of glacial acetic acid. »The liquid was heated to boiling and then left to cool. The liquid was filtered through a drum centrifuge and the filtrate collected and stored. To the filtrate was added 680 g (1.5 pounds) of cetylpyridinium chloride followed by a stirring time of 30 minutes. The liquid was allowed to stand for 16 hours. It formed a precipitate. The above was decanted and the precipitate collected by continuous centrifugation in a Sharples centrifuge. The collected precipitate was in 19 liters (5 gallons) of 0.5N sodium hydroxide. 38 liters (10 gallons) of methanol were added and followed by a room temperature of "12 hours" at room temperature. The precipitate formed was again collected by continuous centrifugation and washed with 19 liters (5 gallons) of methanol. " 2 gallons) of distilled water, the pH was adjusted to 7 _f 0 with glacial acetic acid, and 113 g (1/4 pound) of sodium chloride was added followed by stirring, giving 15 l (4 lbs.

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Methanol) and stirred for 15 minutes. After a standing time of 12 hours at room temperature, a precipitate had formed which was collected by centrifugation. The precipitate was dried under vacuum. The analysis showed that the precipitate was essentially CSA. The material confirmed an extension of the plasma thrombus formation time from 6 to 12 hours after administration to rabbits, as described by the Chandler loop method, of over 80 %,

From the eleventh day of treatment four tablets per day were given in four equal doses for a subsequent period of 13 weeks. "

The treatment duration was 14 weeks, with patients being assessed every 2 weeks during the first six weeks and then monthly during the experimental treatment period. However, prior to the treatment period, assessments were made according to the following scheme:

The first pre-treatment assessment was six weeks prior to the initiation of the treatment period, and patients were further evaluated at two-week intervals in the pre-treatment period.

In all assessments, the general condition of the patient was noted. The clinical severity of claudication and the condition of the skin were classified as mild, moderate or poor. The individual maximum distance of each patient was recorded exactly in meters and any symptoms were noted.

For each assessment, the following additional clinical and laboratory measurements were taken

1. Segmental plethysomogram of one or both lower legs,

2. Micromanomeirische measurement of the arterial blood pressure of the foot,

3. time for onset of calf pain at normal walking speed,

4 "The following subjective measurements of the clinical response:

Number of bouts of calf pain while walking during the day preceding the assessment visit.

Calf pain, measured on a visual analog scale.

Number of periods with calf pain at rest in the week preceding the assessment visit.

Assessment of change based on a visual-n analog scale.

Type and frequency of side effects.

In the following experiment, all subjects had clinically proven arterial occlusive perlveskulitis associated with temporary glaudicatio and signs of impaired arterial blood flow to one or both lower legs. For all subjects, clinical status and disease progression remained relatively constant over the previous six months. None of the patients received anticoagulation therapy or thrombolytic therapy for the previous six months. In addition, people who took vasodilator medications had received them in the previous six to eight months in a constant amount.

.40

People with other symptoms that limited their activity, such as heart disease, were excluded, as well as those with diabetes mellitus and those with vascular surgery on the abdominal aorta, the iliac arteries, superficial and deep femoral arteries, and the terminal branches of the same the lower legs have been made viaren.

A preliminary determination of the following clinical data has been made; Age, weight, height, smoking habits, presence of varicose veins, calf pain at rest and after walking. Patients were categorized by age, sex and cigarette smoking status and randomly assigned to a control group (placebo) or active treatment group (the "active" CSA was prepared according to the above example).

The active test substance was in the form of tablets containing 500 mg of "active" CSA. Placebo tablets were identical in shape, color and consistency to the active tablets, and according to the classification, patients were given randomly for administration of either the active substance or the active ingredient Placebo selected, following sequence was followed?

For the first five days of treatment: 12 tablets daily in four equal doses of three tablets each.

From the sixth to the tenth day of treatment: eight tablets daily in four equal doses of two tablets each.

5. Laboratory investigations:

(a) Hematology: hemoglobin concentration, PCV, MCHC, WBC count and difference count, platelet count, reticulocyte count, and ESR.

(b) plasma binding levels of fibrinogen.

7427

(c) Serum binding levels of cholesterol, liproproteins and triglycerides.

(d) blood coagulation mechanism; Plasma thrombolysis time, euglobulin-LyeiQZieit and platelets ^ aggregation time.

41 patients were involved in the study, that is, twenty-two in the group with active CSA and nineteen in the control placebo group.

All patients had bilateral disease.

The clinical, laboratory and coagulation values of the 41 patients determined in the experiment are shown in Table I. "

Table 1j Clinical and laboratory values of two groups of patients after the experiments: averages - S.Ξ.Μ.

Number of cases Age (years) Cigarette smoker {% of group) % Overweight by size Distance (meters) Calf pain at rest (frequency per week)

Calf pain (visually analogous / mms) 45.6 - 4.5

Fibrinogen (g / l) Plasma clot clotting time {% normal) Buglobulin lysis time (min) Platelet aggregation time (s) Cephalin time (s) Cholesterol (mg / dl) "S""Particle (mg / dl) Maximum calf blood flow (ml / ml) 100 ml tissue / minute)

The difference between the groups is not significant at the 0.05 level.

"CSA" group Placebo- 61 19 0.94 group ί , 7 22 73 _p ,8th 62 ± 1.09 55.9 36 4.7 77.3 11.4 ± 2.0 31.8 50.1 ± 5.7 55.4 ± 3.9 3.37 "Α * 0.13 11.8 ~ 1.9 103 ¹ 1.88 45.6 ± 4.5 330 t 4.24 3.49 ~ 0.16 641 i 4.2 101 ± 1.52 41.3 0.41 327 ^± 5.23 297 ,; 3.48 639 ± 3.2 539 ' 3 i 5.6 40.1 ± 0.34 19.4 1.8 295.1 - 2.8 ± 535.7 ± 6.1 20.8 ± 1.91

The two groups are comparable in terms of age distribution, gestational age, cigar smoke ratings, and their assessment of subjective and objective measures of clinical response.

The improvement in the distance traveled by each of the groups is shown in Table 2

Table 2s Improvement of the route

Therapy: Number of the distance in meters (mean * SEM);

patients:

Pre-trial Week: 0 Week: 2 Week; 4 weeks 6 weeks; 10 weeks; 14

^If CSA "

22 55.4 * 3.9 52.6 * 4.1 53.1 * 3 * 8 78.0 * 5.7 ^ 81, 2 * 4.9 ³ⁱ 86 _? 7 * 5 _f 1 ² 89.3 * 6, 2 ^ä

Placebo 19 55.9 * 4.7 53.8-4.3 54.5 * 4.9 49.3 * 4.5 ^ 49.6 ± 5 ₉ O ^S 50.3 * 4.9 48.7 * 5.1

Significant improvement in "CSA"groups; t _{in (J} = 3.98, P <0.01

  Number of patients with improved distance

Therapy; Total-

cases: week? 0 Week: 2 Week: 4 Week: β Week: 10 Y / oche: 14

"CSA" 22 2 (9.1 %) 6 (27.3 %) 11 (50.0 %) 14 (63.6 %) 15 (68.2 %) 14 (63.6 %) placebo 19 2 ( 10.5 ^) 3 (15.8 %) 2 (10.5%) 4 (21.1 #) 3 (15.8 %) 4 (21.1 9δ)

& Z & & J35 »

- H

In the fourth treatment group, there was a significant improvement in the exercise range in the group receiving chondroitin sodium sulphate (t = -, = $ 3.98 P <0.01). This improvement was also maintained in the following ten weeks of the experiment. There was no significant improvement in exercise in the placebo-administered group.

Table 2 also shows the number of patients who had an improvement in the distance. 3ine would be arbitrary definition; Untreated patient did not show any extension of distance or a minimal improvement (eg an initial 50 m claudication distance was increased to 65 aa). "Survived" patients showed a significant lengthening of walking distance (eg 50 m at the beginning were 80). .. 90 meters extended). From the reproduced in Table 2, it can be seen that the "improved patients" a direct extension of the distance in the fourth, sixth and tenth V / o before DER treatment period showed indicating a progressive improvement.

Table 3 shows changes in the blood circulation (maximum calf flow response) of these two groups of patients.

Table 3% Changes in blood flow (maximum calf bloodstream reaction) in the two patient groups

Max. Blood flow in the calf (ml / 100 ml tissue / minute) Therapy Pre-trial Week: 0 Week: 4 v / o before s 10 Week:

20.8 * 1.91 19.3 * 1.76 27.4 * 1.56 * 35.6 * 1.81 * 34.9 * 1.90 * Placebo 19.4-1.82 20.4- 1.90 18.6 ± 1.73 21.3 * 1.52 20.1 * 1.87

Significant increase in mean blood flow in the "GSA" group P <0.01

It will be noted that in the fourth week of treatment there was a significant increase in average blood flow in the chondroitin sodium sulfate group present at week 10 and week 14 of the trial.

Sine assessment of patients on a change in the thickness of calf pain, as measured by a visual-analogue scale, is included in Table 4.

• Table 4: Assessment of patients on a change in the thickness of calf pain using the iO-cm ~ line: a visual-analog scale

therapy

Num. Of the patients

"CSA" placebo value of t

22 19

Mean - DS Calf Pain (cm) Week 0 Week 2 Week 4 Week 6 Week 10 Week

3,83 * 1,68 4, O1 ± 1,77 4,16 * 1,80 6,44 ^± 1 _i 71 5 _i 63 ^± 2 _J 44 6,10 ^ 2,18 3,76 ± 1y.57 4 , 27 ^± 2, t4 4.38-1.52 4.16-2.03 3.94 ^ 1.97 3.18Ϊ1.88

xnd, •• CSA «group. , _β _ _{Q> 34} _ _{O> 63} . _5F10

Reaction to void 0

Significant improvement only in the "CSA" group P <0.01;

- 2,

-3.86

P <0.001.

Only in the "CSA" group was there a significant improvement, which first became visible in week 6 of the treatment period. Laboratory and coagulation vectors of the two treatment groups determined in the experiment are summarized in the table.

Table 5: Laboratory and coagulation values of the two treatment groups during the experiment (mean * SD)

"GSA" Group (N = 22) Placebo Grup. (N = 19)

Week 4 Week 10 Week 14 Week 4 Week 10 Week

Fibrinogen (g / l) 3.97 * 0.71 3.71 * 0.50 3.48 * 0 * 47 * 3.50 * 0.50 3.52 * 0.55 3.34 * 0.57

Thrombin clotting,,,

time {% normal) 99,7 * 5,32 102,8 * 9,80 104,5-10,76 97 _f 2 * 7,19 102 * 11,10 99,3 * 8,9

Euglobin Lysis Time.

(s) 326.8 * 14.03 331.1 * 16.07 329.2 * 13.64 334.4 * 7.52 327.6 * 8.15 328.5 * 15.24

Bloodsuckle Aggre-. ,. .

gation period (s) 631.5- 7.63 638.5 * 11.46 640.1 * 10.84 646 * 13.33 638.9 * 12.34 637.5 * 13.69

Cephalin time (s) 40.5 * 4 ₉ 8 40.5 * 2.6 40.1 * 2.9 39 ₉ 5 * 3.7 40.5 * 3.0 41.1 * 2.6

Cholesterol (mg / dl) 293.1 * 13.6 290.8 * 14.4 277.5 * 17 # 7 ** 291.0 * 21 _s 3 297.7 -11.9 288.3 * 18.9

^is S "particle (mg / dl) 532.6 * 18.6 538.9 * 20.9 536.5 * 19.7 533.0 * 17.1 537 * 14 ₉ 4 538.0 * 16.1

Significant decrease from baseline: ^s 0.01 <P <0.02; ^S3E P <0.01.

Q - ta -

There were also registered significant reductions in average serum cholesterol Konzentra.tionen and the average fibrinogen levels in the ⁿ CSA "~ group in the 14th IVoehe relative to baseline. For all others, made during the experimental measurements were not other significant changes have been identified.

"Active" CSA tablets gave statistically significantly better results in patients with intermittent claudication than placebo, with a significant improvement in travel distance in those patients who had received "active" CSA. Oral administration of "active" CSA was well tolerated and did not cause any significant side effects.

In the experiment, oral administration of "active" CSA was not accompanied by any significant changes in plasma thrombin clotting time and platelet aggregation time.

Bs, in turn, is not able to track the potential course of action of "active" CSA, but the significant reduction in plaema fibrinogen levels and serum cholesterol levels may be the most significant.

It has also been decided to conduct a controlled trial to assess the efficacy of "active" CSA produced as above in preventing deep vein thrombosis after surgery.

140 patients undergoing major gynecological surgery or similar general surgical procedures were examined. Patients on whom leg or hip surgery had been performed were excluded from the trial. ,

2 /

The following clinical data were recorded? Age, weight, size, hospitalization before surgery, preoperative hemoglobin level Eauchgewohnheiten, presence of varicose Yenen on clinical examination, history of venous · 'thromboembolism, type of surgery and whether äex surgery for benign or malignant disease was made. Patients were categorized by age, gender and cigarette smoking status and were randomly assigned to a control (placebo) or active treatment group. All patients received the usual hospital physiotherapy before and after

were

Surgery and / as soon as possible after surgery ambulatory

treated "

The treatment group received orally 10 g of "active" CSA on the two days preceding surgery and then 5 g of "active" CSA (in single doses) daily for seven days after surgery or until the patient was discharged from the hospital.

The platelets were measured with a Colter thrombometer. The decrease in platelet factor III was determined by the method of Spaet and Cintron, the fibrinogen by the method of Ratnoff and Menzie and by Slektroimnmnodiffusion. Antithrombin III according to Howie's method, inter alia, the factor VIII by a step-step method of Brecken and Ratnoff, the euglobulin-lysis time by the method of KiLsson and Dlow, the serum fibrin-related antigen (FR-antigen) after the Method of Merskey et al., The thrombin time measured by the method of Aylward, et al., And platelet aggregation was measured for adenosine diphosphate, collagen, and adrenaline using the aggregometer method of Bourne and Cross.

All patients were isotopic before surgery

125 Examination of the legs using Kakkar's I-pibrinogen technique. A regular postoperative scan was taken on the first, third and sixth day if, not a significantly high count was achieved in making daily scans. The radioactive count at various points of the leg was considered a direct percentage- "

Record of heart rate recorded. It was assumed that patients had a deep vein thrombosis when an increase of 20 % in the same place was observed on two different days or between two adjacent sites, assuming that this increase lasted longer than 24 hours *.

There were 70 patients in the placebo group and 70 in the active treatment group. The clinical and coagulation values of the 140 patients prior to surgery are listed in Table 6. The two groups were similar in age distribution, gender equality and types of operations performed. The occurrence of the various factors indicating the development of deep vein thrombosis was similar in both groups, "

Table 6: Pre-operative clinical and coagulation values of the groups of patients studied

Narcotic cases Active group placebo-Grumve

Number of balls Age (years)

Cigarette smoker {% of group) Pre-operative stay (days) % Overweight to tall

Presence of varicose veins {% of group) Malignant disease {% of group) Fibrinogen (g / l) Factor VII (% of normal) Euglobul'in lysis time (min) Serum FR antigen (mg / l) Plasma Thrombin clotting time (% of normal)

32 32 2 0.10 52 ί 3 51 - , 5 7 35 37 1 23 4 i 1 ± 4 .1 1-2,5 15 ,8th 5 25 18 25 6.25 6 4 · 3.03 ^έ O ₉ H 2.99 4 116 ί 6 114 -S 312 i 21 326 ί 7 ± 1 8th ± 106-4 104

Cases AkJ: ίy e Gr upjp e P1 β c e b ο - Gr u ρ ρ e

38 44 38 3 , 5 42 i β ί 12 13 13 2 14 0 , 09 2 ± 0.5 ί 5 10 12 3 11 2.96 10 0 2.94 ί 0.08 125 ± 7 123  ± 8 296 ± 1 298 ± 15 7 ί 1 8th ί ι 102 ί 3 103 ± 4 ί

Number of cases Age (years) Female Palls

Cigarette smoker (% of group) Pre-operative stay (days) % Overweight by size Presence of varicose veins Fibrinogen (g / l) Factor VII {% of normal) Buglobulin ™ Lysis time (min) Serum FR antigen (mg / l)

Plasma thrombin coagulation »time (% of normal)

Table 7 shows the general incidence of deep vein thrombosis in the placebo and treatment groups. Fifteen of the twenty-five patients with deep vein thrombosis in the placebo group and six 6ex eleven patients with deep vein thrombosis in the treatment group had positive bilateral leg scans. This is a considerable difference (£ 0.02). Considering the type of surgery in the two groups of patients, there is a considerable reduction in the incidence of deep vein thrombosis in the treated group, especially in those patients on whom oophorectomy has been performed. The occurrence of deep vein thrombosis in both groups is shown in Table 8, depending on the type of surgery.

Table 7: Onset of deep vein thrombosis in both groups of patients

Groups Qty. with low-Nr. With

, Palen veins thrombosis bilateral

Gynecological:

Control 32 12 (37.5 %) 8

Treatment 32 5 (15.6 55) 3

I o

groups

General surgery: control treatment

Total patients

Num. of the cases

38 38

140

Num. with low-Nr. with venous thrombosis bilateral

thrombosis

13 (34.2 %) 6 (15.8 %)

36 (25.6 %)

7 3

21

Table 8s Occurrence of deep vein thrombosis in both groups of patients depending on the operation:

surgery

Hysterectomy abdominalis

Hysterectomy abdominalis and oophorectomy

Bilateral oophorectomy

Pelvic surgery

General surgical laparotomy mastectomy adrenalectomy miscellaneous

Plac ebo Group % treated Grutme:

Αώ.ζ. (%) Qty. (%) Qty. (%) Qty. (%) of patients with low-grade patients with low- grade venous thrombosis thrombosis

15 (50 %) 5 (15.6 %) 16 (50 %) 2 (6.3 %)

9 (28.1 %) 4 (12.5 %) 10 ($ 31.2 5) 2 (6.3 %)

5 (15.6 %) 2 (6.3 %) 4 (12.5 %) 1 (3.1 %)

2 (6.3 %) 1 (3.1 %) 2 (6.3 %) 0 (0)

16 (42.1 %) 8 (21.1 %) 14 (36.8? 5) 2 (5.3 ^)

(23.7 %) 4 (10.5 ^) 10 (26.3 %) 2 (5.3%)

(26.3 %) 1 (2.6 %) S (23.7 ίί) 1 (2.6 %)

(7.9 ^) 0 (0) 5 (13.2%) 1 (2.6 ^)

Excessive blood loss during or after surgery was still a problem in treated patients in patients treated with control.

The platelet aggregation time was markedly increased in patients after active CSA, and in the case of the blood count there was no difference in the two groups and no significant change in the persistence of the persistence.

The introduction of the radioactive fibrinogen assay has demonstrated the extent of the problem encountered by postoperative deep vein thrombosis, and the correlation with the venography has confirmed that this test provides an accurate and rapid method for detecting the occurrence and location of thrombosis in patients after the surgery is.

In general surgical patients thrombi usually begin in the deep sinuses of the calf, and if thrombosis is limited to these yens and the deep tibiavens, the risk of pulmonary embolism is low. The expansion of the thrombus into the poplar veins and the lower pemoral veins increases the risk of pulmonary embolism. It is therefore of paramount importance to prevent postoperative deep vein thrombosis whenever possible and to limit expansion if a calf thrombus develops ".

These results indicate that the presence of postoperative deep vein thrombosis can be reduced by "active" CSA, as revealed by the radioactive fibrinogen test in general surgical patients and gynecological patients. * Ss did not show any complications from the administration of the drug. Tests of platelet function and the effects of various drugs in vitro need not reflect the in vivo situation. Nevertheless, such experiments serve as monitors and have confirmed that active treatment has been performed on patients. From the results presented, it is not possible to demonstrate the probable course of the effect of "active" CSA, although it is also possible that inhibition of platelet

zb

Aggregation and prolongation of the plasma thrombin clotting time play a role. It should be noted, however, that elevated levels of plasma pibrinogen have also been reduced by "active" CSA, and this observation may have been of great importance for the beneficial effect of "active" CSA.

The invention described in detail is intended to be limited only by the legal scope of the following claims.

Claims (10)

A process for the preparation of active chondroitin sulphate A or active chondroitin sulphate C or a mixture thereof, characterized in that an aqueous solution of the chondroitin sulphate (s) is prepared from a corresponding starting material, the solution being a complexing agent for the formation of a precipitate, which is a water-insoluble complex of chondroitin sulphate (s) and the complexing agent, is added, and the chondroitin sulphate (s) is (are) recovered from the complex 2 «method according to item 1, characterized in that it is the starting material to mammalian tissue. 3 «Method according to item 2, characterized in that the starting material is bovine-falciparous cartilage. 4 «method according to item 1, characterized in that the starting material is fish tissue. 5 «method according to any one of the preceding points, characterized in that the starting material is degreased before the formation of the solution. 6 »Method according to one of the preceding points, characterized in that the starting material is ground before the formation of the solution. 7 »The method of any of the above items, characterized in that _f that the starting material is digested with an enzyme for removal of protein. 21.7.1981 58 661 18 8. A process according to any one of the preceding claims, characterized in that the complexing agent is a quaternary ammonium salt or an anionic ironing resin. A method according to any one of the preceding claims, characterized in that the complex is split therefrom by a base or hypertonic saline solution to recover the chondroitin sulfate (s) thereof. 10, "Water-insoluble complex of chondroitin sulfate A and / or chondroitin sulfate C and a complexing agent, ge. , characterized in that it is produced by a method according to points 1 to 8 «.