Ａ。交換講座，第２日，４月１３日（土），午後

≪土肥記念国際交換講座講演≫

１ろ.ろ0～1 4.ろ０

33

座長　佐　野　栄　春

Dohi　Memorial　International　Exchange　Lectureship

Gustav Asboe-Hansen, M. D.

　　Professor and Direc tor of

　　Dermato logy, University of

　　Copenhagen, Copenhagon,

　　Denmark

EXPERIMENTAL　SEARCH　FOR　CONNECTIVE-TISSUE

ＡＣＴ工VE　DRUGS.

　　　　　　The systemic mesenchymoses (Syn.　collagenoses.

collagen diseases) are, and have been for ａ series of

years/　the object of Intensive research in our labora-

tories. They may have ａ different etiology and patho-

genesis. However, cells as well as extracellular sub-

stances are involved in all disease processes.　Ａ１１

members of the group affect mesenchymal tissue in the

extended sense.　　　　　　Reticuloendothelial elements.

blood, bone marrow and bone. may participate actively

in the disease processes・

　　　　　　The disorders are of inflammatory nature, and

all phases of inflammation, as they are known from

　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　ofman

granulation tissue or from the development/from the fetal

stage up to old age are represented. Systemic lupus

34 Ａ，交換講座，第２日，４月１３日（土），午後

erythematosus is an acute inflammatory disease charac-

terized by exudation. Scleroderma Is an Inflammatory

disorder as well. but it is characterized by fibrosis

○ヱ‘"healing”。

　　　　　Inflanmiation Is characterized by increased vascular　　　　　-

permeability, maorophage motiltty and phagocytic activity,

an increased number and activity of granulocytes, especially

eoslnophlles and basophlles. as well as lymphocytes, plasma

cells,　fibroblasts and mast cells。

　　　　　AnInterference with the normal activity of one or

several ０ｆthese elements may impair connective-tissue

growth, regeneration and repair. Histamine, a mast-cell

product, may create an Increased vascular permeability.

and ａ tissue edema, Edema exists for ａ very short time.

or as long ａ｀ｓthere １Ｓan intracellular reserve of acid

glycosaminoglycans (GAG).　These GAG, also products of

mast cells, are released when the immediate surroundings

of the cells become watery, changing the tissue water into

ａ mucinous gel. Hyaluronic acid has ａ strong water bind-

ing capacity. and dermatan sulfate is important for colla-

gen deposition to take place。

　　　　　Fibroblasts produce collagen, and agents Influencing

fIbroblasts interfere with collagen synthesis。

　　　　　Realizing that, in scleroderma, there is an intensi-

fied production of collagen and also of acid glycosamino-

glycans we planned to search for substances Inhlbitintj the

biosynthesis of these substances。

　　　　　Collagen biosynthesis pa5;ses the following steps:

ｈ　Rlbosomal biosynthesis of ｐｒりtocollagen which Is ａ

proline- and lyslnerich precursor polypeptide.

ム　Hydroxylatlon of certain proline and lysine residues

of protocollagen to hydroxyproline and hydroxylys ine by

protocollagen-prollne and protocollagen‘lysine hydroxylases,

respectively.　Cofactors required are Fe"*""*"ions, a-keto-

glutarate, ascorbic acid and atmospheric Ｏ２．

Ａ。交換講座，第２日，４月１３日（土），午後

3. Galactosylation of certain hydroxylysine residues to

galactosyl-o-hydroxylvsine by collagen-galactosyl-transfe-

rase. Cofactors required are Ｍｎ十十ions and uridine-diphos-

phate galactose (DDPGal).

£Glucosylation of certain galactosyl-o-hydroxylysine

residues by collagen-glucosyl-transferase. Cofactors re-

quired are Mn and uridlne-diphosphate glucose (UDPGlu).

steps ３ and 4 (the glycosylatlon steps) are essential for

the extrusion of collagen.

　　　　　ｍ

hydroxylated macromolecules ． Onhydroxylated or unglyco-

sylated macromolecules are not extruded from the cells.

The abnormal macromolecules synthesized and the lack of

their extrusion are believed to slow down the synthesis

of protocollagen in ribosomes.　工njectlon ”１ｎoVo" or Into

organ cultures　　　　　　　　　　　　　　　　　　　ofazetidine-

carboxyllc acid, a prollne analogue, which is incorporated

into the precursor polypeptide instead of proline, inhibits

the hydroxylation and, consequently, the glycosylation

steps.　　　　　　　　　　　　　　　　　’　　　　　　　　　　　　　　｀

　　　　　Substances capable of lnhibitinn the incorporation

of prollne and lysine or of chelating Fe and/or Ｍｎφ＋

ions should be expected to hamper collagen biosynthesis.

　　　　　工ｎour search for druなきpossessing the capacity ｔ０，

１ｎsome way or other, Inhibit the Inflammatory process

which proceeds to fibrosis or sclerosis, we planned to

test ａ greaﾓﾆ variety of substances for their influence on

the synthesis of bone or skin collagen and acid GAG In

tissue culture.

　　　　　Adrenocortlcal sterofds.

　　　　　Much GxperimentaJ. information points to the fact

that adrenal steroids Inhibit the exudative collagen

diseases and the inflammatory processes leading to sclero-

derma as well. Nevertheless, adrenal steroids are agreed

３５

36 Ａ。交換講座，第２日，４月１３日（土），午後

by most clinicians to be relatively poor drugs for sclero-

derma,　insofar as usually ｎ０lasting improvement, thoucrh

plenty of side-effects, result from the treatment。 My

personal experience is, that the effect of adrenal steroids

may be beneficial If thev are used in the active, pro-

gressive phases of the disease process only.　As might

be expected, they have no chance in burnt-out cases, ０ｒ

in phases without inflammatory activity。

　　　　　　　１!ＺΞ２!!extract or L-thyroxine has been used for

scleroderrna through many years,　Until recently, the

rationale of this treatment was lacking ０ｒrather vaguely

formulated. Thyrotropic hormone is ａ growth-stimulating

pituitary factor. It stimulates the formation of mucins.

１．:e.丿acidglycosaminoglYcans/ in the tissues. and in the

presence of growth hormone also collagen is laid down. The

nyxedema produced by thyrQtrot?１ｃhormone (possessing fat-

mobilizing and mucotropic activities) may be reduced by　　－

thyroid hormone. Because sclerodermic tissue seems to be

growth-stimulated, thyroid hormone should be expected to

be an Ideal agent to counteract and hamper scleroderma,

However, L-thyroxine in sufficient closesIs generally badly

tolerated by the sclerodermlc patient who often has myo-

cardlal rouclnosls and fibrosis. In contrast to L-thvroxine,

D-thyroxlne does not increase the metabolic rate; although

In hlgh> dosage, some １･ｓtransformed into the L-lsomer.

D-thyroxlne counteracts thyrotropin release from the pituitary

as much or even stronger than the L-isomer. Therefore,

theoretically, D-thyroxine is the drug of choice. In

fact, it has been shown by us to have the capacity of re-

pressing the sclerodermlc inflammation or regeneration.

It has also been shown by us not to inhibit collagen

biosynthesis directly. only to act by the hormonal feed

-back mechanism mentioned above。

　　　　　　　Severalstudies have been performed on the effect

of chelating agents on the biosynthesis of hydroxyproline.

Ａ。交換講座，第２日，４月１３日（土），午後

　Only　α≪' -dipyrldyl had hitherto been studied in parallel

　experiments on the biosynthesis of both amino acids charac-

　teristic of collagen。

　　　　　　　Particularly active In the following experimentation

　was Dr. Nelly Blumenjcrantz。

　　　　　　　Inaddition to the traditionally known chelators.

　some substances of more ｏｚ;less coinmon medical use, known

　to have certain chelating properties, were included in

　the studies. From the 'beginning.　the purpose of the studies

　was to find inhibitors of collagen and GAG biosynthesis and,

　thus, drugs which might be useful １ｎ the treatment of sclero-

　derma。

　　　　　　　Tibiae or skin from 10-day-old chick embryos were

　dissected under the microscope.　Tissues were then pre

-incubated in ａ medium containing glucose, inorganic salts

　and phosphate buffer for ｌ hour at 37°C. The pre-1ncubation

　was continued for ３０ minutes in the presence of the sub-

　stances assayed. Tissues were then incubated with ５戸Ｃ１

　of (■'■''Oli-proline ｏｒ(１４Ｃ)Ｌ－１ｙｓｉｎｅｆｏｒ２ hours at 37°C

　１ｎparallel experiments. At the end of the incubation period

　the tibiae were homogenized, and the homogenates were dia-

　lysed exhaustively against running tan water。

　　　　　　　Inorder ｔｏａｇｓａｙ(１４Ｃ)ｈｙｄｒｏｘｙｐｒ０１１ｎｅor total　十

　(１４Ｃ)ｈｙｄｒｏｘｙｌｙｓｉｎｅ，ａｌｉｑｕｏｔＳof the dialysed homogenates

　were submitted to acid hydrolysis with ６ Ｎ HCl at 12O''c

　overnight. HCl was then evaporated under reduced pressure・

　(■"■^OHypro was assayed according to Juva & Prockop.　(１４Ｃ)

　Hylys was assayed according to Blumenkrantz & Prockop In

　the following way.　Total (１４Ｃ)Ｈｙｌｙｓwas assayed on an

　aliquot of the acld-hydrolysed sample. Unglycosylated

　(^*C)Hylyg was determined on an unhydrolysed allqu。ｔ．

　Glycosylated ( OHylys was determined by the difference

　between total ｉｎｄｕｎｇｌｙｃｏｓｙｌａｔｅｄ(１４Ｃ)Ｈｙｌｙｓ．　Totaluptakes

　ｏｆ(１４Ｃ)Ｐｒｏ　ａｎｄ(１４Ｃ)Ｌｙｓｗｅｒｅdetermined on allquots of the

37

38 Ａ。交換講座，第２日｡４月１３日（土），午後

（１４Ｃ）ｐｒｏand ｒ１１４Ｃ）Ｌｙｓlabelled undialysable samples, ｉｅ‾

spectively。

　　　　　　　？ｈｅeffect of various doses of different chelators

was studied in relation to uptake of （１４Ｃ）Ｌｙｓand (■'■''oPro

and their hydroxylatlon as well as on the glycosylation of

（１４Ｃ）Ｈｙｌｙｓ●　　　　　　　　　　　　　　　　　　　　　　　　　。

　　　　　　　Xhe results were calculated as dpm per bone pet

per hour of incubation and were expressed as percentage of

control values ． Controls were run without addition of the

ohelators。

　　　　　　　Based upon the effect of the chelators on　　1) total

■"■^Cuptake,　2) biosynthesis of hydroxvproline and hydrりXV-

lysine, and 3) glycosylation of hydroxylysine,　they could

be classified In ３ main groups (Table　）．

Groupユ･The main effect of QCOt'-dipyridyl and 8-hydroxy-

quinollne was an inhibition of hydroxylation of （１４Ｃ）Ｔ｀ｒｏ

ａｎｄ（１４Ｃ）Ｌｙｓ．　Glycosylation was correspondingly inhibited.

Groりr> ２７１　　Inhibition of (■'■^C)Pro ａｎｄ（１４Ｃ）ＬｙｓｉｎｃｏｌｒＰｇｒａｌｔｔｏｌｎ

in relation to the concentration of the chelators in the

medium was observed. Their effect on the hydroxylatlon and

the glycosylation steps was even stronger.　1,10 phenanthro-

line, Na-diethYldithlocarbaraa万te, chlorpromazine, hydralazlne.

procainamide and tetracycline chloride belong to this group.

Group 3し　　eomprlses Ｄ一犬penicillamlne, N-acetyl-D,L-penicilia-

mine and EDTA.　D-penlclllamlne and N-acetyl-D,L-penlcilla-

mine Inhibited the incorporation ｏｆ（１４Ｃ）ｐｒｏａｎｄ（１４Ｃ）Ｌｙｓ

and their hydroxylation in relation to the dose, when this

was above ５ ｍ Ｍ。

　　　　　　　The results show that the various chelators do not

affect incorporation and hydroxvlation of (■'■^c)Pro ａｎｄ（１４Ｃ）

Lys to exactly the same extent. The differences observed

may be related to the fact that. besides their metal se-

questering capacity, they may act by other mechanisms as

well, e.c』;_ competition for a common carrier svstem　でｏｆ

Ａ。交換講座，第２日，４月１３日（土），午後

transport, o×ido-reduction mechanisms・ etc. No chelator

was capable of inhibiting glycosylation of hydroxylyslne

only。

　　　　　　　Glyoosylationof hydroxylysine followed the same

pattern as that of the hydroxylatlon of lysine。

　　　　　　　Thedifferences in the uptake and hydroxylation りｆ

　(１を)Proand (１４Ｃ)Ｌｙｓunder the effect of various chelators

stress the need of determining both ami no acids in ccllagen

‘research。

　　　　　　　Itis worth noticing that substances of common

medical use as hydralazine, procainamide and chlorproma‘

zine affected collagen biosynthesis to ａ considerable GX-

tent.　Hydralazlnc also affected the blosyntheses of

ｇｉｖｅＯ－and/or mucoproteins as appears from the decreased

incorporation of ( C)D-glucosamine into undlalysable

material。

　　　　　　　Hydralazlnehas been reported to be capable of

chelatlng Fe'*"''and Mn Ions. Decreased turnover ｏｆＭｎ＋＋

has been reported In one patient suffering from ａ hydra-

lasine syndrome and in ７ patients with rheumatoid ar-

thritis.　Hydralazlne can also bind　OC-ketoglutarate,

another cofactor required for protocollagen hvdroxylation.

The decreased collagen biosynthesis effected by hydralazine

Is well explained by the mentioned ｃｈｅ!ating and binding

effects。

　　　　　　　Theeffect of various ami no acids on collagen

biosynthesis was also studied on organ cultures of chick

embryo tibiae and skin. Since various ami no acids are

chelators for various metal ions, the question arises if

their influence on collagen biosynthesis is due to metal

chelation, or If, by acting ０ｎspecific carrier systems.

they might affect transport mechanisms and, consequently,

the build-up of the protoco1lagen molecule as well.

39

40Ａ。交換講座，第２日，４月1 3 H (土），午後

　　　　　　　Ifthey act as chelating agents, an effect on the

hydroxylatlon and glycosylatlon steps of collagen synthesis

might be expected. ０ｎthe other hand, it １Ｓwell known

that Incorporation of amlno acids Into proteins is de-

pendent on transport processes。

　　　　　　　Bydifferent mechanisms. other ami no acids than

proline and lyaine might act as factors limiting the in-

corporation of both or one precursor into collagen.　As

stated by Tsurufuji, various factors such as cell membrane

permeability and the capacity ｆｏｒ･the cellular accumulation.

production and interconversion of ami no acids mav be

Important for collagen biosynthesis。

　　　　　　　Ourinterest in mechanisms for the biological

regulation of collag≪n biosynthesis, including the possi-

bility of repression of pathological processes loading to

increased collagen formation, made us study the effect of

different natural and synthetic amino acids on the syn-

thesis of collagen.　Competitive inhibition studies for

the uptake ｏｆ（１４Ｃ）Ｐｒｏand ( C)Lys were conducted in the

presence of such　　　　　　　　　　　　　　　　　　amino acids.

Hydroxylatlon of both to (^*C)Hypro and (■'■^C)Hvlys,and

glycosylation of the latter, were also studied. Organ

cultures of 10-day-old chick embryo tibiae and skin were

used as an experimental model。

　　　　　　　？ｈｅtechnique was thessame as used for chelating

agents. In the external medium, all amlno acids occurred

at an inttial concentration of ５ｍＭ，while controls were

run without addition of ami no acids。

　　　　　　　(Table)？henon-radioactive amino acids utilized

for competition studies were L-isomers unless specified

differently.　Uniformly labelled (^^C)L-lyslne ａｎｄ（１４Ｃ）

L-prollne were used.

Ａ。交換講座，第２日，４月１３日（土），午後

　　　　　　　Ａreduction of the incorporation of either labelled

amlno acid by ａ second ami no acid was considered indicative

of ａ common carrier system.

　The results (Table) indicated that, in different　　-

ways, the amino acids tested influenced the uptake of

(１４Ｃ)Ｐｒｏand (*C)Lys

and their hydroxylation. The ami no

acids could be classified Into three groups, according to

their effects on the uptake of the two precursor ami no

acids. They are as follows 。

　　　　　　　Group1.　These amino acids showed competitive In-

ｈｉｂ!tion of ( C)Pro uptake. Hydroxylatlon ｏｆ(１４Ｃ)Ｐｒｏ

generally followed the same pattern of incorporation. The

uptake and hydroxylation of (１４Ｃ)Ｌｙｓａｎｄglycosylation of

(１４Ｃ〉Hylyswere not affected, although cysteine seemed to

have an inhibitory effect on the formation ｏｆ(１４Ｃ)Ｈｙｌｙｓ。

　　　　　　　Group２ヶDecreased Incorporation and hydroxylation

ｏｆ(１４Ｃ)Ｌｙｓｗａｓobserved under the effect of arginlne,

ornithine, and pipecollc acid. Arginine did not affect

the uptake and hydroxylatlon of ( C)Pro, whereas ornithine

ｄｉｄ。

　　　　　　　GrouΣ?i3. These amlno十acids did not affect the up-

take of ｅｉｔｈｅｒ(１４Ｃ)Ｐｒｏor (･'■^OLys。

　　　　　　　？ｈｅeffect of various doses of L-glutamlne on the

incorporation of (■'"^OPro and its hydroxvlation is presented

in Fig.　　Consistent decreases ０ｆthe total uptake of

( C)Pro, and(f(OHypro formation, were observed in

10-day-old chick embryo tibiae, with levels ranging ｆｒｏ竹1

0.35 to 1.07 mM. Ａ similar decrease of incorporation of

(１４Ｃ)Ｐｒｏand Its hydroxylation was observed when ８－and

12-day-old chick embryo tibiae were incubated in the pre-

sence of similar levels of glutamlne. －　　　　　。

　　　　　　　Ourfinding of selective inhibition of (１４Ｃ)Ｐｒｏ

ｏｒ(１４Ｃ)Ｌｙｓby the first and second groups of amino acids

４１

４２ Ａ。交換講座，第2 B, 4月１３日（土），午後

indicates the possibility of biosynthesis, at least experi-

mental, of ａ collagen with ａ normal hvdroxyproline and ａ

reduced hydroxylyslne content and vice versa。

　　　　　　　Ourresults suggest that, besides the chelatlng

effect of amino acids, they may Interfere with the bio-

synthesis of collagen and other proteins by limiting compe-

titive mechanisms or dilution of the ｒり０１０ｆ precursors 。

　　　　　　　工ｔis noteworthy that glutamine is ａ constituent ■

of most media used for culture of manMnallan cells and

has been extensively recognized as ａ component, which is

essential for growth and differentiation of several cell

types.　Many workers interested In the synthesis of colla-

gen in tissue culture have added L-glutamine to the culture

medium at, according to our data, sufficient concentration

to decrease ( C)Pro incorporation, and have. nevertheless.

measured the formation of (１４Ｃ)ｈｙｄｒｏｘｙｐｒｏｌｉｎｅas ａ para-

meter of collagen synthesis. The presence of glutamine in

such cultures might induce the synthesis of an anomalous

collagen.　工ｆ normal and glycosylated hydroχvlyslne is

synthesized under these conditions, extrusion of an ab-

normal collagen could occur。

　　　　　　　L-Dopais a drug which has been studied with this

technique, because L-tvroslne and L-phenylalanine turned

out to influence collagen synthesis in different V7ays。

　　　　　　　Whentibiae of chick embrvos were Incubated vrlth

(■･■^OPro in the presence of increasing concentrations of

L-dopa, decreasing amounts of (１４Ｃ)Ｈｙｐｒｏｗｅｒｅsynthesized.

Although less pronounced, a reduced Incorporation ｏｆ｛１4Ｃ･)Pro

was also observed. If, in parallel experiments, tissues

were incubated with (･"■^OLys in the presence of increasing

concentrations of L-dopa, decreasing amounts of (■'■^OHylvs

were synthesized, while the incorporation of the precursor

(･'■''OLyswas not Inhibited. Glycosylation of (１４Ｃ)Ｈｙｌｙｓ

was decreased parallel to the hydroxylation ｏｆ（１４Ｃ)Ｌリｓ

Ａ。交換講座，第２日，４月１３日（土），午後

(Table). No significant effect of L-dopa on the incorpora-

tion ｏｆ（１４Ｃ）９１ｕｃｏｓａｍｉｎｅｉｎｔｏglyco- and/or mucoproteins

was observed at the dosages used (Table)。

　　　　The finding that the biosvnthesis of both （１４Ｃ）Ｈｖｐｒｏ

ａｎｄ（１４Ｃ）ＨｙｌｙＳ１Ｓmore vigorously decreased than the in-

corporation of their precursor amino acids suggests that

an underhydroxylated collagen is synthesized in the presence

of L-dopa,　This effect could be due to ａ ohelation of

Fe ions,　one of the cofactors required for the hvdroxv‘

latlon step。

　　　　Special attention was paid to ３ drugs which can

produce the lupold syndrome　(LS)。

　　　　皿

inhibits the biosynthesis of collagen and glyco- and/or

mucoprotelns. Besides, chlorpromazine has been found ｔ６

induce the lupoid syndrome and give rise to photoxic and

photoallerglc dermatoses. Recent experiments showed that.

１ｎ vitro, chlorpromaz ine forms complexes with DNA, ＲＮ入，

hyaluronic acid and heparln.　Irradiation with ultraviolet

light induced chemical degradation of chlorpromazine.

These findings may explain the antinuclear factors occurrinfj

in blood of patients with chloでproma zlne-induced lupoid

syndrome, and also the chlorpromazine-induced phototoxic

and photoallergic dermatoses.

　　Procainamide　is also ａ drug that is known to be　　-

capable of producing ａ lupoid svndrome. The drug decrnases

the uptake ｏｆ（１４Ｃ）Ｐｒｏａｎｄ（１４Ｃ）Ｌｙｓ，ｒｅｓｔｔｉｃｔｓthe hvdroxv‘

lation of both, glycosylation of hydroxylyslne, and thG

incorporation ｏｆ（

mucoproteln.

Oglucosamine Into glyco- and/or

　　　　　　皿　１ｓ an outstanding member of the group

of lupoid syndrome inducers ． In expert!ηentsparallel to

４３

４４ Ａ。交換講座，第２日，４月１３日（土），午後

the already mentioned. hydralazlne was ａ strong inhibitor

of ａ１１steps of collagen biosynthesis. It inhibited the

hydroxylatlon of Pro and Lys to ａ stronger deqree than

their incorporation. This １Ｓtrue also of the glvconylation

of Hylys。

　　　　　　　Ａdiminished biosynthesis and an underhydroxylatlon

of collagen (protocollagen) are possihilities. which can

explain the hydralazlne effect.　By chelatinq Ｆｅ＋＋and Ｍｎ十十

and by combining with OC-ketoqlutarate, hydralazine elimi-

nates some important co-factors required for hydroxylation

of protocollagen, as well as for ･the qlvcosylation of the

already hydroxylated protocollagen･　Considering the fact

that Ｍｎ＋＋ ions are required in the biosynthesis of glvco-

saminoglycans the decreased incorporation ｏｆ（14

C)gluco-

samlne into glvco- and/or圃ucoproteins may also be related

to the Ｍｎ十十-chelatlngeffect of hydralazlne。

　　　　　　　Westudied the hvdralazine-induced lupoid syndrome

in more detail. The condition preferably occurs in sub-

jects who are slow acetylators of the drug.　Under ｅχperl-

mental hydralazine treatment of ａ patient VJith geneΓalized

scleroderma of the acrosclerosls type, a lupold syndrome

developed. This patient was followed up since the start

of the treatment. during the lupus crisis and the treat-

ment of It, as well as after his relief and cure from this

severe disorder. He VMS studiec! with biochenical and

Immunological methods。・２２natients with qeneralized

scleroderma, 12 with localized sclf?roderma, 20 with

systemic lupus erythematosus, 1 with pseudoxanthoma elasttcum.

６０ｎchlorpromazine treatment for mental disease, one

patient under recovery from ａ lupus syndrome Induced bv

treatment for ａ heart disease with procainamide, and １２

clinically normal subjects with negative ANF, who were ’

not taking any drugs, served as controls 。

Ａ。交換講座，第２日，４月１３日（土），午後

　　　　　　　Whenserum of the LS patient was mixed with ａ hydra-

lazine-DNAヽcomplex, a filamentous and membranous precipl-

tate developed during the crisis and the following ４Ｔηlonths,

while no precipitate was formed with any of the control

sera.　No precipitate developed when the serum was mixed

with hydralazlne-denatured DNA complex.　Inactivated

serum plus hydralazine-DNA gave no precipitate。

　　　　　　　IncreasedIgM and IgG were found in the solublllzed

precipitate of patient serum and the hydralazine-DNA complex

after remova！ of DNA with protamlne sulphate. In skin

biopsies obtained during the crisis, IgM and small quan-

　　ｌｔｌ,ヒlesof complement were demonstrated by the direct

lmmunofluoresoence technique.　Bv imreunodiffusion the

presence of precipitating antibodies against DNA was shown。

　　　　　　　Lowurinary values of sodium,potassium, calcium.

phosphate and urea were found during the crisis, v/hile ａ

steep increase in the ｕｒ！naryoutput of acid glycosamino-

glycans　with　no significant changes in collagen meta-

bolltes was　noticed。

　　　　　　　Inview of the fact that the pathology of lupus

er ythema tos us Is characterized by acute exudative in-

flammatlon, and scleroderma by chronic fibrotic "healing"

phenomena, it Is noteworthy that three substances which

can produce an １.e.-like syndrome can inhibit collagen

synthesis.　In our clinic, the same substances are now

on clinical trial as therapeutics for scieroderma。

　　　　　　　Besidesthe three substances. the other collagen

synthesis inhibitors mentioned are being tried on-Patients.

It Is too early to evaluate the clinical results.

４５