JPH1029942A - Therapeutic agent for chronic arthrorheumatism - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a new type therapeutic agent for chronic arthrorheumatism, free from adverse effects by using a specific prostaglandin, especially in a form of fat emulsion as an active ingredient. SOLUTION: This therapeutic agent contains prostaglandin E1 as an active ingredient. Targeting therapy is made possible and excellent anti rheumatics is obtained by emulsifying prostaglandin E1 in a form of fat emulsion. Further, action of the prostaglandin E1 is synergically enhanced by combinedly using the prostaglandin E1 with a phosphodiesterase inhibitor such as pentoxifylline in a low concentration. The prostaglandin E1-containing fat emulsion preferably contains prostaglandin E1 [in an amount of 0.001-0.01% (W/V)], plant oil, at least one kind of fat emulsion base (in an amount of 5-20%) selected from a 6-12C middle chain fatty acid triglyceride and 6-18C fatty acid di- and monoglyceride, an emulsifier of phospholipid (in an amount of 0.6-2.4%) and an isotonizing agent of glycerin and water and has <=1μm average particle diameter.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

[0001] The present invention relates to a therapeutic agent for rheumatoid arthritis, and more particularly to prostaglandin E1 (PGE).
The present invention relates to a therapeutic agent for rheumatoid arthritis comprising 1) as an active ingredient. Furthermore, the present invention also relates to a therapeutic agent for rheumatoid arthritis, comprising a combination of PGE1 and a phosphodiesterase inhibitor having an action of synergistically enhancing the anti-rheumatic action of PGE1.

[0002]

BACKGROUND OF THE INVENTION The analgesic and antipyretic effect of aspirin is controlled by the prostaglandin E2 through the cyclooxygenase (COX) inhibitory action of the arachidonic acid metabolic pathway.
It has been clarified that suppression of (PGE2) production has caused COX inhibitors to be classified as anti-inflammatory drugs because they suppress acute inflammatory models such as carrageenan edema. Since then, PGE2 has been considered as one of the proinflammatory substances of acute inflammation, and various types of COX inhibitors that specifically suppress the production of PGE2 have been developed as non-steroidal anti-inflammatory drugs (NSAIDs). It has been used to treat many inflammatory diseases. In rheumatoid arthritis (RA: sometimes simply referred to as RA hereinafter),
Since PGE2 is produced locally in arthritis, NSAIDs have been administered for the purpose of suppressing the production. However, although NSAIDs reduce morning stiffness and arthralgia in patients with rheumatoid arthritis, they do not have the effect of suppressing joint destruction or deformation progression due to arthritis, and do not improve the indicators of arthritis such as erythema and CRP. That is, the NSAID basically has no antirheumatic effect.

[0003] At present, the main therapeutic agents for rheumatoid arthritis are auranofin, which is an injection or oral preparation which is considered to have an anti-rheumatic effect, furthermore, methotrexate (MTX), salazopyrine, and bucillamine. However, the efficacy of such antirheumatic drugs is less than 60%, and serious side effects are frequently observed. Therefore, at present, there is a strong demand for the development of an antirheumatic drug that is more effective and has no problems such as side effects.

On the other hand, PGE2 has been found to have various anti-immune and anti-inflammatory effects in vitro.
That is, T cell proliferation and interleukin-2 (IL
-2) Suppression of production, activation and proliferation of B cells, activation of monocytes / macrophages, and suppression of inflammatory cytokine production. However, whether or not PGE2 exerts an antirheumatic effect has not been examined since 1971. That is, in 1971, Zuri of New York University
er et al. reported that a very large amount of prostaglandin E1 (PGE
1) is administered and its effectiveness is reported. In this report, a 500 μg subcutaneous injection of PGE1 was given twice daily,
Although administered daily, transient shocks, impaired consciousness and the like were observed after the administration, and it was difficult to administer such a large amount in humans. Therefore, it was completely unknown whether PGE1 itself had an anti-rheumatic effect to suppress the pathology specific to RA.

On the other hand, in 1983, Mizushima et al. Reported that PGE was contained in small fat particles (lipid microspheres) in a fat emulsion.
Lipo-PGE1 (fat emulsion PGE1) encapsulating (embedding) 1 is being developed. This lipo-PGE1
Was effective for peripheral circulatory disorders such as arteriosclerosis obliterans by administering 10 μg per day. lipo-PGE1 is
By making PGE1 into a fat emulsion, small fat particles are selectively taken up locally in the lesion, and PG encapsulated on the spot
E1 has been shown to exert an effect, and has been used as a PGE1 targeting therapeutic agent in the treatment of various circulatory disorders. However, still lipo-P against RA
Examination of the therapeutic effect of GE1
At present, no attempt has been made even in vivo.

In view of the above situation, the present inventors have been studying the development of an effective therapeutic agent for RA in recent years, and in the course of the study, among prostaglandins, prostaglandin E (PGE) )). That is, in local arthritis of rheumatoid arthritis, PGE is considered to be one of the biological defense response factors for suppressing the pathological state of rheumatism, and among PGEs, PGE1 particularly has a specific therapeutic effect on RA. It was thought that there was. Therefore, from this point, it can be said that administration of an NSAID having no anti-rheumatic effect to RA patients, on the contrary, suppresses PGE production, and may worsen the pathology of RA. Also, if PGE is one of the biological defense response factors for suppressing the disease state of rheumatism, if the sufficient amount of PGE1 can be introduced locally to arthritis by applying the targeting therapy proposed by Mizushima et al. And thought that treatment would be possible.

The present inventors have developed a cell model that efficiently reproduces synovitis of rheumatoid arthritis in vitro, and has already filed a patent application for this point. In this cell culture system, proliferation of synovial cells characteristic of RA, production of inflammatory cytokines, proliferation of transformed fibroblasts, and formation of pannus-like tissue are reproduced. That is, this system is used in an in vitro human synovitis model, that is, in vitro for chronic indirect rheumatism.
It can be said that this is a pathological model in the field.

Accordingly, the present inventors have studied the effects of PGE1 on this in vitro human synovitis model using the cell model proposed by the present inventors in conducting the above-mentioned studies. In addition, it is an animal model of RA II
PGE1 fat emulsion (lip
oPGE1) was administered, and the therapeutic effect based on the targeting therapy was also examined. As a result, PGE1 becomes in
An anti-rheumatic effect was recognized in vitro and in vivo, and it was newly found that a fat emulsion of PGE1 had a more excellent anti-rheumatic effect, thereby completing the present invention.

The various biological activities possessed by PGE1 are mainly due to the cyclic AMP (cy
clicAMP) is known to be exerted by increasing the concentration. Therefore, intracellular cyclic
Under circumstances where AMP levels can be maintained at high levels, P
It is considered that the biological activity of GE1 can be exhibited more effectively. In fact, phosphodiesterase inhibitors such as pentoxifylline are cyclic A
Inhibits MP metabolism and increases intracellular cyclic
It is known that the AMP concentration is maintained for a long time, and that the biological activity is synergistically enhanced when both are administered in combination, and clinically the action of PGE1 on vascular endothelial cells and platelets Has attracted attention, and a combination therapy of both has been applied to improve cerebral circulation and peripheral circulation.

On the other hand, the anti-rheumatic effect of pentoxifylline itself has also been studied, and its large dose (eg, 1,6
(00 mg / day) was reported to be effective in half of rheumatoid arthritis patients. However, in the treatment by such a large amount of administration, side effects such as gastrointestinal symptoms naturally become a problem, and thereafter, it has not been clinically applied. From this point of view, it is impossible to develop a practical therapeutic agent for rheumatoid arthritis based on the antirheumatic effect of pentoxifylline alone. Therefore, the present inventors focused on the phosphodiesterase inhibitory action of pentoxifylline. That is, when pentoxifylline is co-administered with PGE1 at a dose that does not show an anti-rheumatic effect, intracellular
It has been newly confirmed that cyclic AMP can be maintained at a high level, and as a result, the above-mentioned anti-rheumatic effect of PGE1 is synergistically enhanced, thereby completing the present invention.

[0011]

Accordingly, an object of the present invention is to provide an antirheumatic drug which is more effective and has no problems such as side effects.

[0012]

Means for Solving the Problems The object of the present invention to solve such problems is to provide a therapeutic agent for rheumatoid arthritis containing prostaglandin E1 (PGE1) as an active ingredient. Furthermore, the present invention provides, in a specific embodiment thereof, a therapeutic agent for rheumatoid arthritis in which PGE1 is in the form of a fat emulsion.

By the way, a fat emulsion of PGE1 has already been proposed by Mizushima et al. And is used for the treatment of peripheral circulatory disorders such as atherosclerosis obliterans. However, this emulsion has no antirheumatic effect. It was not known. Therefore, another object of the present invention is to provide a PGE1-containing fat emulsion as a therapeutic agent for rheumatoid arthritis. Specifically, (a) prostaglandin E1 (P
GE1): 0.001-0.01% (w / v), (b)
Vegetable oil, at least one fat emulsion base selected from triglycerides of medium chain fatty acids having 6 to 12 carbon atoms and di- and monoglycerides of fatty acids having 6 to 18 carbon atoms: 5 to 20% (w / v), (C) Phospholipid emulsifier: 0.6 to 2.4% (w / v), (d) glycerin isotonic agent, and (e) water, having an average particle size of 1 micron or less A prostaglandin E1-containing fat emulsion as a therapeutic agent for rheumatoid arthritis.

In the present invention, several patent applications have already been filed for the fat emulsion itself in which PGE1 is encapsulated in lipid microspheres (for example, Patent No. 1,289,054). Fat emulsions containing various drugs, for example, anti-inflammatory analgesics such as flurbiprofen and 4-biphenylylacetic acid, various steroids or SOD have been proposed (for example, Japanese Patent No. 1,921,169: 61-44809). The fat emulsion in this case is generally a lipid microsphere (small fat particles) having an average particle size of 1 micron or less.
Wherein the composition comprises at least one fat emulsion selected from the vegetable oils described above, triglycerides of medium-chain fatty acids having 6 to 12 carbon atoms, and di- and monoglycerides of fatty acids having 6 to 18 carbon atoms. It has a basic composition of a base, an emulsifier that is a phospholipid, a tonicity agent that is glycerin, and water, and the mixing ratio of each component is also within the range of each of the above numbers. Therefore, a fat emulsion embedding PGE1 as a therapeutic agent for rheumatoid arthritis of the present invention is also a fat emulsion having such a general constitution, and specific examples of each component are described in the specifications of these preceding patent applications. And the specification is included as part of the present specification.

The prostaglandin E1 (PGE1) -containing fat emulsion as a therapeutic agent for rheumatoid arthritis proposed by the present invention has already been used for the treatment of peripheral circulatory disorders such as obstructive atherosclerosis as described above. lipo-
PGE1 (fatty emulsion PGE1) is preferably used, but is not limited to this. The amount of PGE1 can be variously changed according to the purpose of treatment, and the desired RA can be obtained by adjusting the fat emulsion composition in accordance therewith. It can be a PGE1-containing fat emulsion as a therapeutic agent. That is,
The dose of PGE1 used for the treatment of rheumatoid arthritis in the present invention is preferably determined by the symptoms of rheumatoid arthritis or by its administration means, and is exemplified in the above-mentioned prior patent specification. It goes without saying that specific components can be appropriately combined to prepare a desired PGE1-containing fat emulsion capable of administering such a dose.

Another object of the present invention is to provide an effective dose of prostaglandin E1 (PGE1) and an effect of a phosphodiesterase inhibitor sufficient to synergistically exert the action of PGE1. The present invention provides a therapeutic agent unit for rheumatoid arthritis, characterized in that both dosages are used in combination.

In this case, it has been found that pentoxifylline is particularly preferable as the phosphodiesterase inhibitor co-administered together with the effective dose of PGE1.
Therefore, as another aspect, the present invention provides a phosphodiesterase inhibitor as an enhancer for the anti-rheumatic effect of PGE1, and more preferably, PGE1 containing pentoxifylline as an active ingredient. And an anti-rheumatic effect enhancer. In this case, the form in which the effective dose of PGE1 is administered is not particularly limited as a therapeutic agent unit for rheumatoid arthritis, but it can be administered in the form of the PGE1-containing fat emulsion described above.

On the other hand, when a phosphodiesterase inhibitor represented by pentosquiphyllin is used in combination, a low dose which does not exert an antirheumatic effect by itself is sufficient. The combined use of toxifylline with PGE1 provides a therapeutic agent unit for rheumatoid arthritis which has a strong antirheumatic effect and has few side effects.

The details of the specific anti-rheumatic effect of PGE1, which is an active ingredient of the present invention, and the details of the anti-rheumatic effect in combination administration with pentoxifylline as a phosphodiesterase inhibitor will be described below. The present invention will be described.

I: PGE1 in vitro
Effect : The effect of PGE was examined using an in vitro human synovitis model for evaluating the efficacy of an antirheumatic drug, for which the inventors have already filed a patent application. Methods : Synovial tissue obtained during synovectomy or artificial joint replacement in rheumatoid arthritis patients was minced and 10% F
RPMI-1640 with CS or 5% human AB serum
The cells were cultured in a medium. On the 3rd to 5th days after the start of the culture, the floating cells are collected and subjected to primary mixed culture at a concentration of 1 to 2 × 10 ⁶ / ml. The culture form was observed over time, and the amount of cytokine produced in the culture supernatant was measured. The cytokine was measured using an ELISA kit. The proliferative response is
The MTT assay was used.

Results: (1) It was confirmed that PGE1 suppressed the proliferation of synovial cells. After synovial cells were cultured in basal medium in the presence or absence of various concentrations of PGE1 for 4 to 9 days, MTT
Proliferation response was measured by an assay. Figure 1 shows the results.
As shown. FIG. 1-a is from day 4 and FIG.
Shows the one on the sixth day, and FIG. 1-c shows the one on the ninth day. As is clear from the results in the figure, PGE1 and PGE1
It is understood that both GE2 suppress synovial proliferation in a dose-dependent manner at a concentration of 0.01 μM to 1 μM. On the other hand, in the presence of the PGE synthesis inhibitor indomethacin, synovial growth was conversely enhanced. The result is shown in FIG.

(2) PGE1 is TNFα produced by synovial cells
It was confirmed that production was suppressed. When synovial cells were cultured in a basal medium, TNFα was produced in the culture supernatant. This TNFα production was suppressed in the presence of PGE1 or PGE2 and increased in the presence of indomethacin. These results are shown in FIG.

(3) PGE1 is in vivo by synovial cells
It was confirmed that tissue formation in vitro was suppressed. i
The inhibitory effect of PGE on a human cell model that reproduces synovitis tissue in vitro was examined. That is, in order to semi-quantitatively express the morphological changes after the culture, the scores were given as follows. Cell aggregation and appearance of blank area: 1 point Appearance of extracellular matrix: 2 points Multi-layered cell aggregation in matrix: 3 points Macroscopically visible tissue formation: 4 points Average score of 3 holes in culture dish The value was taken as the organization score.

FIG. 4 shows the state of the morphological change. As shown in the figure, in the control group cultured only in the basal medium, the morphological change progressed after 1.5 weeks, and after 3.5 weeks, a visually observable tissue was formed. This tissue formation is based on PGE1
Alternatively, it was suppressed in the presence of 0.2 μM of PGE2. This inhibitory action of PGE1 was demonstrated by adding PGE1 to 0.001 μM,
The concentration was changed to 0.01 μM, 0.1 μM and 1 μM, and the results of the examination are shown in FIG. As is clear from the results in FIG. 5, it was confirmed that the inhibitory effect of PGE1 was dose-dependent. Also, PGE1 was 0.002 μM,
The concentration was changed to 0.02 μM and 0.2 μM,
FIG. 6 shows the results of examining the inhibitory action of No. 1 together with the effect of indomethacin 0.1 μM. As is clear from the results in the figure, PGE1 suppresses in a dose-dependent manner, whereas indomethacin, a PGE synthesis inhibitor, conversely enhances tissue formation. The results of these (1) to (3) are:
This clearly shows that PGE1 has an anti-rheumatic effect in vitro.

II: Effect of PGE1 in vivo
RESULTS : Type II collagen arthritis was DBA / 1 in a 6 week old male.
It was prepared by a conventional method described in the literature using J mice. Method: First, on the first day, 100 μg of type II collagen was mixed with complete Freund's adjuvant and injected subcutaneously at the tail of the mouse. On day 21, 50 μg of type II collagen was mixed with incomplete Freund's adjuvant and injected subcutaneously. Second
On day 5, the mice were randomly divided into four groups, and group A contained 100 μl (0.5 μg) of commercially available lipo-PGE1 used for treatment of peripheral circulatory disorders such as atherosclerosis obliterans.
In group B, 100 μl of fat emulsion intrafat (lipo-PGE1 carrier) was injected as a control, and in group C, 100 μl of physiological saline was intravenously injected into the tail vein every day for 10 days.

For evaluation of arthritis, the incidence and severity were determined every other day. The incidence rate is represented by (number of affected limbs / number of mouse limbs per group), and the severity is unchanged for each limb .... 0 point 1 finger or multiple fingers 1 point redness and swelling of the whole ... 2 points Intense swelling of the whole ... 3 points with change in joint tonicity ... 4 points The total number of limbs was set to 16 points, and the result was represented by an average score per mouse.

Results: (1) DBA / 1J mice immunized with type II collagen were boosted on day 21 to develop arthritis, and from day 25, 100 μl (0.5 μg) of lipo-PGE1 was added to 10
Intravenous injections were given daily for days. The incidence of arthritis in mice receiving lipo-PGE1 was lower than in mice receiving control fat emulsion or saline. The result is shown in FIG. Arthritis was also less severe. The result is shown in FIG. As is clear from the results of FIG. 7 and FIG. 8, after the treatment with lipo-PGE1 (35 days after immunization), the incidence of arthritis increased and was no different from the other groups (FIG. 7). The severity of arthritis did not increase (FIG. 8). From this point, it is understood that the therapeutic effect of lipo-PGE1 is maintained for at least 10 days after discontinuation of administration.

(2) Forty-two DBA / 1J mice that also developed arthritis with type II collagen were randomly divided into four groups of 12 mice each, and from day 25, 0.3 mg of lipo-PGE1 or methotrexate (MTX). FIG. 9 shows the results of intravenous administration of / kg for two weeks every day. The severity of arthritis was greater than that of the control group by 200 for lipo-PGE1.
It is understood that the value is significantly lower in the μl (1 μg) administration group. In this respect, 50 μl of lipo-PGE1 was used.
(0.25 μg) in the administration group also tended to be less severe. In contrast, no difference was found in the severity of the MTX-administered group from the control group (FIG. 9). From the above results, PGE1 exerts an excellent antirheumatic effect even in vivo, and particularly, lipo as a fat emulsion.
-It is understood that PGE1 shows a superior effect.

III: Cyclic AM in vitro
Efficacy of P-elevating drug : Similar to I described above, the present inventors have already applied for a patent, and have used an in vitro human synovitis model for evaluating the efficacy of antirheumatic drugs.
The effects of AMP-elevating drugs were examined. Methods : Synovial tissue obtained during synovectomy or artificial joint replacement in rheumatoid arthritis patients was minced and 10% F
RPMI-1640 with CS or 5% human AB serum
The cells were cultured in a medium. Three to five days after the start of the culture, the cells that have deviated from the tissue are collected and subjected to primary mixed culture at a concentration of 1 to 2 × 10 ⁶ / ml. The culture form was observed over time, and the amount of cytokine produced in the culture supernatant was measured. The cytokine was measured using an ELISA kit. The proliferative response used the MTT assay.

Results: (1) Dibutylyl-cyclic AMP which is a cell membrane permeable cyclic AMP (hereinafter referred to as db)
-cAMP) was confirmed to suppress the growth of synovial cells as in PGE1. After synovial cells were cultured for 6 days in a basal medium in the presence or absence of various concentrations of db-cAMP or PGE1, the proliferation reaction was measured by MTT assay. The result is shown in FIG. FIG. 10 (a)
Indicates that of db-cAMP, and FIG. 10B shows that of PGE1. As is clear from the results in the figure, db-cAM
It is understood that P inhibits synovial cell proliferation in a dose-dependent manner at a concentration of 1 μM to 100 μM.

(2) db which is a cell membrane permeable cyclic AMP
It was confirmed that -cAMP suppressed the production of TNFα similarly to PGE1. It turns out that TNFα produced in the culture supernatant of synovial cells is suppressed in a dose-dependent manner in the presence of db-cAMP, similarly to PGE1. These results are shown in FIG. FIG. 11A shows the result of db-cAMP, and FIG. 11B shows the result of PGE1.

(3) db which is cyclic AMP permeable to cell membranes
-cAMP is similar to PGE1 in synovial cells in vivo.
It was confirmed that it suppressed in vitro tissue formation. in
In order to semi-quantitatively show the morphological change of synovitis tissue in vitro, the same scoring as described in the examination of I above was performed. In addition, the average value of the scores of the two wells of the culture dish was used as the organization score. FIG. 12 shows the result. As is clear from the results in the figure, in the control group cultured only in the basal medium, the morphological change progressed after one week, and a tissue that could be visually observed was formed two weeks later. This tissue formation is db-cAM
It turns out that it is suppressed in a dose-dependent manner in the presence of P. Judging from the results of (1) to (3) above,
By increasing the intracellular cyclic AMP concentration, PG
It is confirmed that the antirheumatic effect of E1 is reproduced.

IV: Pentrishi in vitro
Anti-rheumatic effect of filin : The anti-rheumatic effect of pentoxifylline was examined in the same manner as in the above method III. Results: (1) It was confirmed that pentoxifylline suppressed the growth of synovial cells at a concentration of 30 μg / ml. After synovial cells were cultured in a basal medium in the presence or absence of various concentrations of pentoxifylline for 6 days, the proliferation reaction was measured by MTT assay. The result is shown in FIG. As is clear from the results in the figure, it is understood that pentoxifylline inhibited the growth of synovial cells at a concentration of 30 μg / ml, but not at a concentration of 3 μg / ml or less.

(2) Pentoxifylline has a concentration of 10 μg /
It was confirmed that the production of TNFα was suppressed by about 30% at a concentration of ml. TNFα produced in the culture supernatant of synovial cells
At a concentration of 10 μg / ml of pentoxifylline of about 3
It was found that the inhibition was 0%, and at a concentration of 3 μg / ml, the inhibition was as small as 20% or less.
These results are shown in FIG.

(3) Pentoxifylline is 3 μg / m
It was confirmed that the concentration of 1 or more suppressed in vitro tissue formation by synovial cells. Inv by synovial cells
The effect of pentoxifylline on tissue formation in vitro was examined in the same manner as in III- (3) above. FIG. 15 shows the average score of tissue formation after 3 weeks of culture. As shown in the figure, pentoxifylline inhibited in vitro tissue formation by synovial cells in a dose-dependent manner, but a concentration of 3 μg / ml or more was required for its significant inhibition. Prove. Judging from the above results (1) to (3), it was shown that the antirheumatic effect of pentoxifylline requires a concentration of at least 3 μg / ml or more.

V: PGE1 in vitro
Synergistic antirheumatic effects of pentriciphyrin : IV above
Test results showed that the concentration at which the anti-rheumatic effect of pentoxifylline was expressed was at least 3 μg / ml or more, so that pentoxifylline was co-administered with PGE1 at a low concentration at which the anti-rheumatic effect was not exhibited In this case, the effect of pentoxifylline on the anti-rheumatic effect of PGE1 was examined. The test method is
The procedure was performed in the same manner as described in III.

Results: (1) It was confirmed that a low concentration of pentoxifylline enhances the inhibitory effect of PGE1 on synovial cell proliferation. After synovial cells were cultured in a basal medium in the presence or absence of pentoxifylline 1 μg / ml and various concentrations of PGE1 for 6 days, the proliferation reaction was measured by MTT assay.
The result is shown in FIG. The abbreviation symbol PO in the figure
F means pentoxifylline and therefore w / oP
OF means in the absence of pentoxifylline, w / POF means in the presence of pentoxifylline (same in the following figures). As is clear from the results in the figure,
1 μg of pentoxifylline showing no antirheumatic effect
At a low concentration of / ml, it was found that the inhibitory effect of PGE1 on synovial cell proliferation was enhanced.

(2) The low concentration of pentoxifylline
It was confirmed that GE1 enhances the effect of suppressing the production of TNFα. TNFα produced in the culture supernatant of synovial cells
It was suppressed by PGE1, but this inhibitory effect was found to be further enhanced in the presence of 1 μg / ml of pentoxifylline. The result is shown in FIG.

(3) The low concentration of pentoxifylline is
It was confirmed that TNFα of GE1 enhances the in vitro inhibitory effect of synovial cells on tissue morphology. III-
In the same manner as in (3), i-cells by synovial cells were obtained by using a combination of low concentration of pentoxifylline and low concentration of PGE1.
The effect on tissue formation in vitro was examined.
FIG. 18 shows the average score of tissue formation after 3 weeks of culture. As shown in the figure, 1 μg of pentoxifylline
/ Ml was hardly suppressed (does not show an antirheumatic effect). At 2 nM concentration of PGE1, it was only slightly suppressed. However, the in vitro tissue formation is remarkably suppressed by the combination of the two, and it is understood that the low concentration of pentoxifylline synergistically enhances the anti-rheumatic effect of PGE1.

[0040]

As described above, the present invention is the first in vitro method for PGE1 which has not been studied.
The anti-rheumatic action in o and in vivo was confirmed, and based on such action, it is specific in that it provides an anti-rheumatic drug containing PGE1 as an active ingredient. In addition, by emulsifying PGE1 in a fat emulsion,
There is an advantage that a targeting therapy can be performed and a better antirheumatic drug is provided. Furthermore, the anti-rheumatic effect of PGE1 is mediated by an increase in intracellular cyclic AMP, and the effect is synergistically enhanced by the combined use of a low concentration of a phosphodiesterase inhibitor such as pentoxifylline. Became clear. Such a therapeutic agent unit for rheumatoid arthritis having a specific synergistic effect provides a completely new therapeutic agent without side effects. Thus, it provides great promise for the treatment of patients with chronic rheumatism for which no effective treatment has been available.

[Brief description of the drawings]

FIG. 1 is a view showing the effect of PGE1 and PGE2 of the present invention on synovial cell proliferation in vitro.
1-a shows the result on the fourth day, 1-b shows the result on the sixth day, 1
-C shows the results on day 9.

FIG. 2 is a graph showing the effects of both PGE1 and indomethacin of the present invention on synovial cell proliferation in vitro.

FIG. 3 shows TNFα of PGE1 and indomethacin of the present invention.
It is a figure which shows the effect of production suppression in vitro.

FIG. 4 shows in vitro of PGE1 synovial cells of the present invention.
FIG. 9 is a diagram showing an effect on tissue formation at o.

FIG. 5 shows in vitro of PGE1 synovial cells of the present invention.
FIG. 9 is a diagram showing the results of observing the effect on tissue formation at o by changing the concentration.

FIG. 6 shows the results of observing the effects of PGE1 and indomethacin of the present invention on tissue formation of synovial cells in vitro at different concentrations.

FIG. 7 shows the effect of lipo-PGE1 of the present invention in v
It is a figure which observes in vivo and shows the result of the incidence rate of the arthritis.

FIG. 8 shows the effect of lipo-PGE1 of the present invention in v
It is a figure which shows the result of the severity of the arthritis observed in vivo.

FIG. 9 is a graph showing the effects of lipo-PGE1 and MTX of the present invention in vivo and the results of the severity of arthritis.

FIG. 10 shows the effect of PGE1 and db-cAMP of the present invention on synovial cell proliferation in vitro. 10-a shows the result of db-cAMP, and 10-b shows the result of PGE.
1 shows the results.

FIG. 11 shows TNF of PGE1 and db-cAMP of the present invention.
FIG. 2 is a view showing the effect of suppressing α production in vitro. 11-a shows the result of db-cAMP, and 11-b shows the result of PGE.
1 shows the results.

FIG. 12 shows the effect of db-cAMP on tissue formation of synovial cells in vitro.

FIG. 13 shows the effect of pentoxifylline on synovial cell proliferation in vitro.

FIG. 14 is a graph showing the effect of pentoxifylline on the inhibition of TNFα production in vitro.

FIG. 15: Pentoxifylline in synovial cells in vivo.
It is a figure which shows the effect with respect to the tissue formation in tro.

FIG. 16: In V against synovial cell proliferation by combination of PGE1 of the present invention and pentoxifylline at low concentration.
It is a figure which shows the effect in vitro.

FIG. 17 shows the in vivo inhibition of TNFα production by the combined use of PGE1 of the present invention and pentoxifylline at a low concentration.
It is a figure which shows the effect in vitro.

FIG. 18 is a graph showing the effect of the combined use of PGE1 and pentoxifylline at low concentrations of the present invention on tissue formation of synovial cells in vitro.

Claims (8)

[Claims] 1. A therapeutic agent for rheumatoid arthritis comprising prostaglandin E1 (PGE1) as an active ingredient. 2. A therapeutic agent for rheumatoid arthritis in which prostaglandin E1 (PGE1) is in the form of a fat emulsion. (A) Prostaglandin E1 (PGE)
1): 0.001 to 0.01% (w / v), (b) selected from vegetable oils, triglycerides of medium-chain fatty acids having 6 to 12 carbon atoms, and di- and monoglycerides of fatty acids having 6 to 18 carbon atoms. At least one fat emulsion base: 5
Emulsifier which is 〜20% (w / v), (c) a phospholipid:
0.6 to 2.4% (w / v), (d) a tonicity agent that is glycerin, and (e) water, and as an agent for treating rheumatoid arthritis having an average particle size of 1 micron or less. Prostaglandin E1-containing fat emulsion. 4. An effective dose of prostaglandin E1 (PGE1) and a phosphodiesterase (Phosphodiesteras) sufficient to express the action of PGE1 synergistically.
e) A therapeutic agent unit for rheumatoid arthritis, wherein both effective doses of the inhibitor are used in combination. 5. A phosphodiesterase (Phosphodiest) used in combination with prostaglandin E1 (PGE1).
5) The therapeutic unit for rheumatoid arthritis according to claim 4, wherein the inhibitor is pentoxifylline. 6. The therapeutic agent for rheumatoid arthritis according to claim 4 or 5, wherein the therapeutic agent for rheumatoid arthritis according to claim 2 or 3 is used as an effective dose of prostaglandin E1 (PGE1). 7. A phosphodiesterase inhibitor as an enhancer for the anti-rheumatic effect of prostaglandin E1 (PGE1). 8. An anti-rheumatic action enhancer of prostaglandin E1 (PGE1) containing pentoxifylline as an active ingredient.