WO1998027990A1

WO1998027990A1 - Methods of elevating amounts of hyaluronan in the human body

Info

Publication number: WO1998027990A1
Application number: PCT/CA1997/000847
Authority: WO
Inventors: Stefan Gustafson
Original assignee: Hyal Pharmaceutical Corp
Current assignee: Hyal Pharmaceutical Corp
Priority date: 1996-12-24
Filing date: 1997-11-12
Publication date: 1998-07-02
Anticipated expiration: 1999-06-24
Also published as: AU4938397A

Abstract

The use of an effective amount of chondroitin sulphate (CS) is provided to cause a human being to provide an elevated therapeutically effective amount of hyaluronan in the human body.

Description

TITLE OF THE INVENTION

Methods of Elevating Amounts of Hyaluronan in the Human Body. FIELD OF THE INVENTION This invention relates to methods of increasing the levels of hyaluronan in the human body to therapeutically effective amounts of hyaluronan in the human body. BACKGROUND OF THE INVENTION

In Canadian Applications 2,164,260 and 2,173,037 I disclosed my new methods of treating diseases and conditions using chondroitin sulphate based on my discovery that while chondroitin sulphate and hyaluronan bind to liver cells and particularly the same receptors of the liver, chondroitin sulphate does not bind with the receptors on, tumours (e.g. metastatic tumours) and other pathological sites (for example, regions of restenosis). This discovery led me to develop my new methods of treatment of diseases and conditions including metastatic tumours, underperfused tissue, diseased tissue, etc. By saturating the receptors on the liver cells by administered chondroitin sulphate (or other GAGS [Glycosaminoglycans] such as dextran sulphate, other than a form of hyaluronan) and subsequently administering the form of hyaluronic acid (hyaluronan) with a medicine and /or therapeutic agent, the subsequently administered amounts of hyaluronan (which transport the medicine and /or therapeutic agent) are picked up, not by the liver whose binding capacity has been substantially filled by the GAG but by other sites capable of binding with forms of hyaluronan having excess unfilled hyaluronan receptors (such as on the metastatic tumours). At the same time, the hyaluronan transports any medicine (or therapeutic agent) to treat the sites in need of treatment (for example, an effective amount of a cytotoxic agent to treat a tumour). As a result I have by the pre- treatment with for example chondroitin sulphate found that less of the form of the transport agent hyaluronan that is to be subsequently administered is required than previously used. I have also found that less of the medicine and /or therapeutic agent is required than was used previously to provide a successful treatment of the disease and /or condition. The amount of the chondroitin sulphate that will saturate the liver cells I found was preferably in the order of at least about 3-5 mg of chondroitin sulphate per kilogram of body weight of the patient. Greater amounts (mg/kg) may be administered to prolong the effect. Because the liver processes the administered and "taken up" chondroitin sulphate rapidly, less chondroitin sulphate is not as good as more as after several hours the liver has processed all the chondroitin sulphate. Thus, prolonged, "blockage" /"down regulating" or "immobilization" of the liver cells is preferred. Where chondroitin sulphate (equivalent dose 1-2 grams/ 70 kg person) is administered, the take-up of even small amounts of hyalyronan by the liver (0.5-1 mg/70 kg person) can be inhibited for an extended period of time by the administration of the chondroitin sulphate. Thus, the hyaluronan is available to transport the medicine and /or therapeutic agent to the site in need of treatment (for example, methotrexate or cisplatin to a tumour, or furosemide to a kidney or other use proposed by the teachings of WO 91/04058 and other documents which were incorporated by reference into Canadian Patent Applications 2,164,260 and 2,173,037). SUMMARY OF THE INVENTION

I have now also discovered that the administration of chondroitin sulphate or other glycosaminoglycan (GAG) such as dextran sulphate, dermatan sulphate, and keratan sulphate (but not heparin) and other molecules with affinity for the liver hyaluronan receptor to the human body such as by intravenous administration or intramuscular or subcutaneous injection or by oral administration in a suitable excipient for example chondroitin sulphate in saline or for oral administration in an excipient suitable to form a tablet, capsule and the like, elevates the amount of hyaluronan in the human body. Thus, by administration of a suitable amount of chondroitin sulphate, the amount of hyaluronan may be elevated from the usual or expected amounts to therapeutically effective amounts of hyaluronan in the human body such as for example, to inhibit or prevent, restenosis (the narrowing of arterial walls) for example, after balloon angioplasty (see Publication No. WO 95/26193 by Hyal Pharmaceutical Corporation) or, inflammatory conditions such as inflammation of joints or to treat or prevent or inhibit other conditions or diseases whose treatment or prevention is aided by the presence in the human body of an excess amount of hyaluronan [more than the usual amount in circulation in the human body (40-140 mg/ml)], to therapeutically effective amounts. For prevention of restenosis, page 53 of WO 95/26193 describes administration of amounts of forms of hyaluronan for therapeutic use as follows: "In view of the results, optimal dosages for administration to humans is {sic} in the order of about 250 mg/70 kg person to 1500 mg/70 kg person with suggested amount being in the order of between about 500 mg to about 1000 mg/70 kg person (at this amount in the order of 15 mg/kg) of the form of hyaluronic acid sodium hyaluronate. The forms can be administered by injection, by subcutaneous administration or by intravenous (sterile) administration from IN. bags."

The amount of chondroitin sulphate (or other receptor binding molecule or GAG such as dextran sulphate, keratan sulphate or dermatan sulphate) may also be combined with amounts of forms of hyaluronan (hyaluronic acid or pharmaceutically acceptable salts thereof) if instant high levels of hyaluronan is desired. The amount of hyaluronan added [such as exemplified in the specification of International Publications No. WO 91/04058, and WO 95/26193 will vary depending on the amount of chondroitin sulphate, (or other agent) used to elevate the amount of hyaluronan in the human body to the therapeutically effective amount (for example to elevate the amount of hyaluronan in the fluid to greater than 20,000 ng/ml (20 μg/ml) for example 1000 μg/ml. Thus, the amount of chondroitin sulphate or other agent (glycosaminoglycan) administered (with or without a form of hyaluronan) to achieve levels of hyaluronan in the human body equivalent to the administration of the exemplified amounts referred at for example page 53 of WO 95/26193 may be suitable. The amounts of chondroitin sulphate (or other agent) (with or without the form of hyaluronan) may be administered in any suitable tolerable form previously known to persons skilled in the art.

Thus, according to an aspect of the invention, I have provided the use of an effective amount of chondroitin sulphate (CS) to cause an elevated therapeutically effective amount of hyaluronan in the human body (for example in the circulation system, vascular system, lymph system, for example in the blood system, proximate the arteries around the heart).

According to another aspect of the invention I have provided the use of an effective amount of glycosaminoglycan (GAG) except heparin to cause an elevated therapeutically effective amount of hyaluronan in the human body. Examples of such GAGS comprise dextran sulphate, dermatan sulphate and keratan sulphate. (While not technically a glycosamino-glycuronoglycan, keratan sulphate and the like will be considered herein to be included in the expression glycosaminoglycan (GAG).

According to another aspect of the invention the use of the amount of the GAG such as chondroitin sulphate, dextran sulphate, keratan sulphate, and dermatan sulphate is sufficient to cause an elevated therapeutically effective amount of hyaluronan in the human body to prevent (inhibit) restenosis.

According to another aspect of the invention, the use of the amount of the GAG such as chondroitin sulphate, dextran sulphate, keratan sulphate, and dermatan sulphate is sufficient to cause an elevated therapeutically effective amount of hyaluronan in the human body to treat inflammatory conditions.

According to another aspect of the invention I have provided a method of elevating the usual or normal amount of hyaluronan expected to be in the human body to a therapeutically effective amount of hyaluronan in the human body, the method comprising administering a therapeutically effective amount of glycosaminoglycan (GAG) (except heparin) to elevate the usual or normal amount of hyaluronan expected to be in the human body to a therapeutically effective amount of hyaluronan in the human body. The glycosaminoglycan may comprises chondroitin sulphate, dextran sulphate, keratan sulphate, and dermatan sulphate. This method may be used to prevent arterial restenosis or to treat an inflammatory condition. According to another aspect of the invention I have provided the use of an effective amount of glycosaminoglycan (GAG) excluding heparin in the manufacture of a pharmaceutical composition for elevating the amount of hylauronan in the human body to an elevated therapeutically effective amount of hyaluronan in the body. According to another aspect of the invention, I have provided a dosage amount of a pharmaceutical composition suitable for systemic administration comprising in a suitable diluent for administration, an amount of a glycosaminoglcan (GAG) [except heparin] exceeding at least about 3 mg/kg of body weight. According to another aspect of the invention, I have also provided methods and uses for sustaining the therapeutically effective amount of the hyaluronan in the human body by administering a further therapeutically effective amount of the GAG after a predetermined time after the earlier administration to elevate and sustain the therapeutically amount of hyaluronan in the human body.

According to another aspect of the invention, hyaluronan may also be added to the human being treated to instantly elevate the amount of hyaluronan in the body. The GAG to follow may comprise chondroitin sulphate, dextran sulphate, dermaton sulphate, and keratan sulphate.

The amount of GAG (e.g. chondroitin sulphate) administered in each dosage is variable over a broad range of amounts. A person skilled in the art will appreciate that the maximum useful amount of the GAG (for example chondroitin sulphate) is that amount which will saturate the receptors with which it can bind. When the receptors are saturated, no further effect can be provided by the GAG. At that time the liver receptors are essentially bound and the GAG (e.g. chondroitin sulphate) can inhibit the uptake of hyaluronan by the lymph nodes further increasing the circulating levels of hyaluronan. As time passes the GAG is metabolized thus permitting the hyaluronan circulating to be taken up by previously blocked or bound receptors. Thus, sustaining dosage amounts of the GAG (e.g.. chondroitin sulphate, dextran sulphate, and keratan sulphate (with or without a form of hyaluronan) may be given to the patient being treated at predetermined intervals to sustain (maintain) the level of the desired therapeutically effective amount of the hyaluronan in the body for example to prevent arterial restenosis or treat inflammatory conditions. Thus, an individual would be administered in each dosage at least about 3 mg of chondroitin sulphate per kilogram of body weight per hour. This administration should therefore provide an effective concentration of hyaluronan in the blood exceeding 200 ng/ml (200 μg/litre) of blood.

Thus, for example chondroitin sulphate may be administered to provide amounts of greater than for example lμg/ml [one microgram per milliliter] of chondroitin sulphate in the bodily fluid. Because the affinity constant for chondroitin sulphate is less than 1 μg/ml (e.g. .5 μg/ml [500 nanograms/ml of blood for example]), the giving of more than about 3 mg/kg of body weight in a suitable dosage form is only effective if given after for example a few hours after the earliest administration to sustain the level of the GAG and thus maintain the amount of the hyaluronan in for example the blood. Preferably the dosing of the GAG is given to maintain medium- high levels of hyaluronan in the bodily fluids. The presence of the hyaluronan is then available to prevent arterial restenosis or treat peripheral inflammation such as inflammation of the joints or to treat or prevent other known conditions where above normal amounts of hyaluronan are required and desirable in the human body.

The invention will now be illustrated with reference to the following discussion of embodiments thereof having regard to the following Figures 1, 2 and 3. BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 illustrates the effect of intravenous chondroitin sulphate on the levels of hyaluronan in rat serum. 0.5 ml of 10 mg/ml chondroitin sulphate in sterile phosphate buffered saline was injected into a tail vein of Sprague Dawley rats. Before the injection, time =0, a blood sample, approximately 0.5 ml, was collected. Blood was then collected from the cannulated vein at 10 and 20 minutes, and a larger blood sample from the aorta at 70 minutes. Serum was prepared and analysed for hyaluronan by the use of an HA-50 kit from Pharmacia & Upjohn, Uppsala, Sweden. Open squares represent a rat receiving also 10 μg HA. Filled squares represent mean values± SD of three rats receiving chondroitin sulphate with only negligible amounts of hyaluronan.

Figure 2 illustrates the effect of intravenous dextran sulphate (DxS, Mw 500 kDa, Pharmacia & Upjohn, Uppsala, Sweden) and phosphate buffered saline (PBS) control. Open squares represent a rat receiving 0.5 ml of a solution of lOOmg DxS/ml PBS. The DxS solution contained no hyaluronan and did not react in the HA-assay. Filled squares represent mean values±SD of three rats receiving only 0.5 ml PBS. (For details of administration see discussion with respect to Figure 1 above.)

Figure 3 illustrates effect of intraperitoneal chondroitin sulphate on the circulating levels of hyaluronan. Three rats were given 0.5 ml of a solution of 50 mg/ml chondroitin sulphate. Before the injection (0 min) a blood sample was collected from the tail vein. At 180 min the rat was killed and blood drawn from the aorta. Serum was prepared and analysed as described in Fig. 1 Values are mean values +SD. Statistical analysis was performed using student T-test analysis with Statworks® on a Macintosh II si computer.

Figure 4 illustrates serum from the initial blood sample (squares), the blood sample taken at 70 minutes after chondroitin sulphate injection (open circles) and urine collected at 70 minutes (filled circles) were subjected to size exclusion chromatography on a 50 ml column of Sephacryl SI 000 and S300 calibrated with hyaluronan standards of known Mw:s. After separation the fractions were analysed for hyaluronan and the total amount of hyaluronan present in the serum and urine calculated from the volume of urine collected and total blood volume estimated to be 6% of the body weight. DETAILED DISCUSSION

I have discovered that a consequence of blocking of chondroitin sulphate (CS)/hyaluronan (HA) liver receptors by chondroitin sulphate is that this will cause an increase in circulating hyaluronan caused by the endogenous influx, mainly via the lymph, to the general circulation. This influx is estimated to be about 40-80 mg/d under normal circumstances. As the liver is responsible for over 90% of the uptake and degradation of circulating hyaluronan, inhibition of liver uptake results in a massive accumulation of hyaluronan in plasma. Thus the inhibition of the degradation of hyaluronan in circulation, for example for one day will result in a concentration of hyaluronan in plasma of about 20 000 ng/ml by the end of the day, to be compared with the normal level of 30-50 ng/ml. As the hyaluronan is continuously being broken down in the circulation, most of the hyaluronan will, after some time, be cleared via the kidneys and lost in the urine.

Lymphatic hyaluronan normally entering the circulation has a very high Mw (in order of several million Da). As the binding of hyaluronan to the receptors is Mw dependent, high Mw polymers are preferentially cleared via chondroitin sulphate (CS)/ hyaluronan (HA) receptors in both liver and lymph nodes. When chondroitin sulphate is given, the rather low Mw (mean above 30 kDa) results in a large part of the chondroitin sulphate not only to bind to liver receptors but also to distribute in interstitial fluid and lymph where it can inhibit the uptake of hyaluronan by the lymph nodes. This will result in increased amounts of high Mw hyaluronan bypassing the nodes and entering the general circulation in comparison to the normal situation. Thus, high doses of chondroitin sulphate will give increased blood levels and urinary excretion of hyaluronan by interfering with uptake and degradation in peripheral tissues also.

That chondroitin sulphate has the above discussed effect is seen in studies on rats that are injected intravenously with 5 mg highly pure chondroitin sulphate (90% 6-sulphate, 10% 4-sulphate). When the serum hyaluronan levels were studied the increase was from 40-140 ng/ml prior to injection to 500-900 ng/ml at 70 min. after injection (Fig. 1). The hyaluronan level increased rapidly and approached a steady state level of about 10 times the initial value at about 70 min after injection (Fig 1). The increase in total plasma hyaluronan was between 3.8 and 8.1 μg at 70 min, and as the chondroitin sulphate preparation contained only 60ng hyaluronan/mg chondroitin sulphate, only 0.3 μg could be attributed to injected hyaluronan. Furthermore, at 70 min. the urine was found to contain 10 000 - 70 000 ng/ml at the 70 min. time point with a total excretion of 3.5-7.5 μg.

When the serum from blood drawn prior to injection and at 70 minutes were subject to size-exclusion chromatography on a calibrated column of Sephacryl®, hyaluronan could be detected over essentially the entire Mw spectrum but most of the material eluted with a mean Mw of about 100 kDa in both cases (Fig 4.) the urinary hyaluronan was of relatively low Mw with a mean Mw of around 10 kDa (Fig 4.)

When 5mg chondroitin sulphate (80% 4-sulphate and 20% 6- sulphate), containing also 2 μg HA/mg CS, was injected intravenously and the blood levels followed at 1, 10, 20 and 70 minutes, it was found that blood levels increased from 1 min. (when it was already high due to the injection 10 μg HA present in the preparation) up to 10 min. and then stayed at a steady state level significantly higher than when hyaluronan was given with only minute amounts of hyaluronan (Fig 1). Thus, chondroitin sulphate can given in combination with hyaluronan to achieve a prolonged high level of hyaluronan without giving high doses of hyaluronan or continuous infusions. It could be given by intravenous as well as by intramuscular or subcutaneous injections. As intramuscular, intraperitoneal, or subcutaneous injection would give a prolonged effect on the hyaluronan levels as the injected material will stay for some time as a depot in the tissues. Intramuscular single injections of 200 mg chondroitin sulphate has been found to give high levels of chondroitin sulphate in serum with a peak concentration about 100 times normal at 90 minutes (Ronca G, and Conte A; Int. J. Clin. Pharmaceol. Res. 13, 27-34). Repeated injections of very high doses and injections at several sites will give increased hyaluronan levels and also prolongation of the effect. The effect of intraperitoneal injection is shown in (Fig 3). As chondroitin sulphate has been shown to reach high levels in blood between 2 and 24 h after oral administration of 0.8 g in humans, oral chondroitin sulphate before and after intravenous hyaluronan should be a convenient way of sustaining a high hyaluronan level e.g. in the treatment of inflammatory conditions or after balloon angioplasty. However, as some chondroitin sulphate will be broken down and desulphated before reaching the bloodstream, oral treatment should be carefully monitored and adjusted in patients and will have to be tested empirically for the effect on serum hyaluronan at several different doses and conditions.

I have achieved equally good results with chondroitin-6- sulphate as well as with chondroitin-4- sulphate. The preparations show great polydispersity with a mean Mw around 30 kDa and 90% of molecules between 10 kDa and 100 kDa. Dermatan sulphate, dextran sulphate (DxS) and other related polysachharides can also be used. However, synthetic polysaccharides such as dextran sulphate (DxS), cannot be preferred due to toxicity and immunological reactions. Natural polysaccharides that are constantly present in higher animals, such as chondroitin sulphate are preferred. A high Mw is preferred because of better binding to chondroitin sulphate (CS) /hyaluronan (HA) receptors. However, to achieve the effect also on the lymph nodes, as discussed above, a medium high Mw (20-40 kDa) is preferred as these molecules will distribute also in the interstitial fluid and lymph system. Of course the reverse is also true, namely that hyaluronan therapy (giving of hyaluronan) will affect the metabolism of chondroitin sulphate and other polysaccharides, but as these generally have lower Mw.s their accumulations in plasma will probably not be so dramatic as for hyaluronan. As many changes can be made to the embodiments without departing from the scope of the invention, it is intended that all material contained herein be interpreted as illustrative of the invention and not in a limiting sense.

Claims

THE EMBODIMENTS OF THE INVENTION FROM WHICH AN EXCLUSIVE PROPERTY OF PRIVILEGE IS CLAIMED ARE AS FOLLOWS:

1. The use of an effective amount of chondroitin sulphate (CS) to cause a human being to provide an elevated therapeutically effective amount of hyaluronan in the human body.

2. The use of Claim 1 wherein the therapeutically effective amount of hyaluronan is in the blood.

3. The use of Claim 1 wherein the therapeutically effective amount of hyaluronan is present in the arteries proximate the heart.

4. The use of Claim 1, 2, or 3 wherein the amount of hyaluronan exceeds about 200 ng/ml (200 μg/1) of blood.

5. The use of an effective amount of a glycosaminoglycan (GAG) except heparin to cause a human being to provide an elevated therapeutically effective amount of hyaluronan to be present in the human body.

6. The use of Claim 5 wherein the GAG is selected from dextran sulphate, dermatan sulphate and keratan sulphate.

7. The use of Claim 5 wherein the therapeutically effective amount of hyaluronan is in the blood.

8. The use of Claim 6 wherein the therapeutically effective amount of hyaluronan is in the blood.

9. The use of Claim 5 wherein the therapeutically effective amount of hyaluronan is present in the arteries proximate the heart.

10. The use of Claim 6 wherein the therapeutically effective amount of hyaluronan is present in the arteries proximate the heart.

11. The use of Claim 5, 6, 7, 8, 9, or 10 wherein the amount of hyaluronan exceeds about 200 μg/1 of blood.

12. The use of Claim 1, 2, or 3 wherein the effective amount of chondroitin sulphate is in an amount of at least about 3 mg/kg of human body weight sufficient to give a concentration of at least about 1 μg/ml of bodily fluid.

13. The use of Claim 5, 6, or 7 wherein the effective amount of the GAG is an amount of chondroitin sulphate is an amount of at least about 3 mg/kg of human body weight sufficient to give a concentration of at least about 1 μg/ml of bodily fluid.

14. The use of Claim 8, 9, or 10 wherein the effective amount of the GAG is an amount of chondroitin sulphate is an amount of at least about 3 mg/kg of human body weight sufficient to give a concentration of at least about 1 μg/ml of bodily fluid.

15. The use of Claim 1, 2, or 3 wherein the amount of chondroitin sulphate is sufficient to cause an elevated therapeutically effective amount of hyaluronan to prevent (inhibit) restenosis.

16. The use of Claim 5, 6, or 7 wherein the amount of GAG is sufficient to cause an elevated therapeutically effective amount of hyaluronan to prevent (inhibit) restenosis.

17. The use of Claim 8, 9, or 10 wherein the amount of GAG is sufficient to cause an elevated therapeutically effective amount of hyaluronan to prevent (inhibit) restenosis.

18. A method of elevating the usual or normal amount of hyaluronan expected to be in the human body to a therapeutically effective amount of hyaluronan in the human body, the method comprising administering a therapeutically effective amount of chondroitin sulphate (CS) to elevate the amount of hyaluronan in the body to a therapeutically effective amount of hyaluronan in the body.

19. The method of Claim 18 wherein the therapeutically effective amount of hyaluronan is in the blood.

20. The method of Claim 18 wherein the therapeutically effective amount of hyaluronan is present in the arteries proximate the heart.

21. The method of Claim 18, 19, or 20 wherein the amount of hyaluronan exceeds about 200 ng/ml of blood.

22. A method of elevating the usual or normal amount of hyaluronan expected to be in the human body to a therapeutically effective amount of hyaluronan in the human body, the method comprising administering a therapeutically effective amount of glycosaminoglycan (GAG) (except heparin) to elevate the usual or normal amount of hyaluronan expected to be in the human body to a therapeutically effective amount of hyaluronan in the human body.

23. The method of Claim 22 wherein the GAG is selected from dextran sulphate, dermatan sulphate and keratan sulphate.

24. The method of Claim 22 wherein the therapeutically effective amount of hyaluronan is in the blood.

25. The method of Claim 23 wherein the therapeutically effective amount of hyaluronan is in the blood.

26. The method of Claim 22 wherein the therapeutically effective amount of hyaluronan is present in the arteries proximate the heart.

27. The method of Claim 23 wherein the therapeutically effective amount of hyaluronan is present in the arteries proximate the heart.

28. The method of Claim 22, 23, 24, 25, 26, or 27 wherein the amount of hyaluronan exceeds about 200 ng/ml of blood.

29. The method of Claim 18, 19, 20, or 21 wherein the effective amount of chondroitin sulphate is an amount of at least about 3 mg/kg of human body weight sufficient to give a concentration of at least about 1 μg/ml of bodily fluid.

30. The method of Claim 22, 23, 24, 25, 26, or 27 wherein the effective amount of the GAG is an amount of chondroitin sulphate is an amount of at least about

3 mg/kg of human body weight sufficient to give a concentration of at least about 1 μg/ml of bodily fluid.

31. The method of Claim 18, 19, 20, or 21 wherein the amount of chondroitin sulphate is sufficient to cause an elevated therapeutically effective amount of hyaluronan to prevent (inhibit) restenosis.

32. The method of Claim 22, 23, 24, 25, 26, or 27 wherein the amount of GAG is sufficient to cause an elevated therapeutically effective amount of hyaluronan to prevent (inhibit) restenosis.

33. The use of an effective amount of chondroitin sulphate (CS) in the manufacture of a pharmaceutical composition for elevating the amount of hylauronan in the human body to an elevated therapeutically effective amount of hyaluronan in the body.

34. The use of Claim 33 wherein the therapeutically effective amount of hyaluronan is in the blood.

35. The use of Claim 33 wherein the therapeutically effective amount of hyaluronan is present in the arteries proximate the heart.

36. The use of Claim 33, 34, or 35 wherein the amount of hyaluronan exceeds about 200 ng/ml of blood.

37. The use of an effective amount of a glycosaminoglycan (GAG) excluding heparin in the manufacture of a pharmaceutical composition for elevating the amount of hylauronan in the human body to an elevated therapeutically effective amount of hyaluronan in the body.

38. The use of Claim 37 wherein the GAG is selected from dextran sulphate, dermatan sulphate and keratan sulphate.

39. The use of Claim 37 wherein the therapeutically effective amount of hyaluronan is in the blood.

40. The use of Claim 38 wherein the therapeutically effective amount of hyaluronan is in the blood.

41. The use of Claim 37 wherein the therapeutically effective amount of hyaluronan is present in the arteries proximate the heart.

42. The use of Claim 38 wherein the therapeutically effective amount of hyaluronan is present in the arteries proximate the heart.

43. The use of Claim 37, 38, 39, 40, 41, or 42 wherein the amount of hyaluronan exceeds about 200 ng/ml of blood.

44. The use of Claim 33, 34, or 35 wherein the effective amount of chondroitin sulphate is an amount of at least about 3 mg/kg of human body weight sufficient to give a concentration of at least about 1 μg/ml of bodily fluid.

45. The use of Claim 37, 38, 39, 40, 41, or 42 wherein the effective amount of chondroitin sulphate is an amount of at least about 3 mg/kg of human body weight sufficient to give a concentration of at least about 1 μg/ml of bodily fluid.

46. The use of Claim 33, 34, or 35 wherein the amount of chondroitin sulphate is sufficient to cause an elevated therapeutically effective amount of hyaluronan to prevent (inhibit) restenosis.

47. The use of Claim 37, 38, 39, 40, 41, or 42 wherein the amount of GAG is sufficient to cause an elevated therapeutically effective amount of hyaluronan to prevent (inhibit) restenosis.

48. A dosage amount of a pharmaceutical composition suitable for systemic administration comprising in a suitable diluent for administration an amount of a glycosaminoglycan (GAG) [except heparin] exceeding at least about 3 mg/kg of body weight.

49. The dosage of Claim 48 wherein the GAG is chondroitin sulphate.

50. The dosage of Claim 48 wherein the GAG is dextran sulphate.

51. The dosage of Claim 48 wherein the GAG is keratan sulphate or dermatan sulphate.

52. The dosage amount of Claim 48, 50, and 52, wherein the dosage amount is in the form for intravenous administration.

53. The dosage amount of Claim 48, 50, 52, wherein the dosage amount is in injectable form ( e.g. subcutaneous or intramuscular).

54. The dosage amount of claim 53 wherein the dosage amount is in injectable form for subcutaneous administration.

55. The dosage amount of claim 53 wherein the dosage amount is in injectable form for intramuscular administration.

56 The method of claim 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 or 32 further comprising sustaining the therapeutically effective amount of hyaluronan in the body of a human by administering a further therapeutically effective amount of a GAG (other than heparin) after a predetermined time after the earlier administration to elevate and sustain the therapeutically amount of the hyaluronan in the human body.

57. The dosage amount of claim 53 wherein the dosage amount is in oral form for oral administration.

58. The method of claims 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, or 32 wherein hyaluronan is also added to the human body.

59. The use of any previous use claim wherein hyaluronan is also added to the human body.