20
On insulin-like actions of morphine and certain morphine derivatives.‘ BY Gunnar Ahlgren, Uppsala. (With 9 flgores in the text.) The intimate connection between the many functions of the living cells and the biological oxidation processes makes the question of the influence of drugs on cell oxidations an important task for research. Our modern methods for the study of cell and tissue oxidation havc also opened a new and extensive field for experimental pharmacology. In this paper are presented the results of a series of investigations in which the action of morphine, some other opium alkaloids, and certain morphine derivatives on the oxidation processes in frogs’ inuscles has been studied. They have been investigated mainly by the aid of Thunberg’s methylene blue method, but partly also with the micro- respirometric technique of the same author. It was during work on the influence of insulin on tissue metabolism that the question of the action of morphine and related substances became of interest. It was found that insulin exerts a typical action on tissue oxidation; an action which is in all probability intimately related to the primary physiological function of insulin. It was possible to demonstrate that insulin, in cooperation with glucose and a certain agent, glueornutin, present in the cells, strongly accelerates the oxidation rate in the tissues under certain conditions2 [Ahlgren (I)]. Received for publication October 20th, 1929. 2 It must be emphasized that insulin does not primarily increase tissue oxidation. The accelerating action is a secondary one. I refer to my analysis of this problem in a previous paper. - Ahlgren, 1. Skandinav. Archiv. LVIII. 11

On insulin-like actions of morphine and certain morphine derivatives

Embed Size (px)

Citation preview

Page 1: On insulin-like actions of morphine and certain morphine derivatives

On insulin-like actions of morphine and certain morphine derivatives.‘

BY Gunnar Ahlgren,

Uppsala.

(With 9 flgores in the text.)

The intimate connection between the many functions of the living cells and the biological oxidation processes makes the question of the influence of drugs on cell oxidations an important task for research. Our modern methods for the study of cell and tissue oxidation havc also opened a new and extensive field for experimental pharmacology.

In this paper are presented the results of a series of investigations in which the action of morphine, some other opium alkaloids, and certain morphine derivatives on the oxidation processes in frogs’ inuscles has been studied. They have been investigated mainly by the aid of Thunberg’s methylene blue method, but partly also with the micro- respirometric technique of the same author.

It was during work on the influence of insulin on tissue metabolism that the question of the action of morphine and related substances became of interest.

It was found that insulin exerts a typical action on tissue oxidation; an action which is in all probability intimately related t o the primary physiological function of insulin. It was possible t o demonstrate that insulin, in cooperation with glucose and a certain agent, glueornutin, present in the cells, strongly accelerates the oxidation rate in the tissues under certain conditions2 [Ahlgren (I)].

Received for publication October 20th, 1929. 2 It must be emphasized that insulin does not primarily increase tissue

oxidation. The accelerating action is a secondary one. I refer to my analysis of this problem in a previous paper. - Ahlgren, 1.

Skandinav. Archiv. LVIII. 11

Page 2: On insulin-like actions of morphine and certain morphine derivatives

154 GUKN.ZR AHLGRES :

Nost of the experinieiits were performed with the niethylene blue method ; but the main results were also confirmed by microrespironieter experiments [Brahme (2). Ahlgreii (3)] and with Lipsrh i tz ’ dinitro- bensol method [Ahlgren (I), p. 1661. The main results have been coii- firmed by Macleod and Markowitz (4)’ B l i s (5), Gigon (6), Hol- boll (7) and von Eu le r jr . (8), all of whom used the methylene blue method. as well as by Wolf (9), working in Kin te r s t e in ’ s laboratory with microrespironietric technique. The conflicting observations of sonic other investigators are dealt with in a previous discussion [Ahlgren (lj, p. 16411 - where also some indirect confirmations of the experimental data are referred to.

Regarding the theory and the technique of the methylene blue method, reference should be made t o the full description given in in?; monograph, or that given in Abderhalden’s Handbuch [Ahlgren (1) resp. (lo)] or Thunberg’s presentation in Oppenheimer’s ,,Die Fermente“ (11); as t o the microrespirometric technique, the papers of Brahme (2) and Ahlgren (3) should be consulted.

Methylene blue experiments.

The main features of the methylene blue technique are as follows: In a vacuum tube of the Thunberg pattern 200 nig. finely cut

muscle (or other) tissue from an animal, just previously killed, are suspended in 1 cc. of a solution of methylene blue and K,HPO,. The tube is then evacuated and put into a waterbath. Occasionally the tube is shaken by hand, and the time required for the complete decoloration of the dye is measured.

The rate of decoloration of the dye stuff in such a tube will be in- fluenced by insulin in various ways, according t o the circumstauws. The following observations have been made in experiments with insulin. Their interpretation and the case of certain exceptions have been already dealt with [Ahlgren (l)]:

1. Addition of insulin alone causes - above a certain concentration - a retardation of the decoloration (Experiment 1, curve J) . The retarding action increases with the insulin concentration.

glucose, which without insulin does not influence the rate of reduction, accelerates the decoloration, even when the insulin concentration is as low as 10-l6. The acceleration increases as the insulin concentration is ra,ised until an optimum point

2. Addition of insulin plus

1 Later experiences of certain experimental difficulties that may be some- times encountered will be dealt with in a special paper.

Page 3: On insulin-like actions of morphine and certain morphine derivatives

O X INSULIIV-LIKE ACTIOSS OF MORPHINE, &C. 155

(at about 10-l2) is reached. Then the acceleration diminishes with further rise in the concentration of insulin until when it has reached about 10-8 there is no detectable action. Even higher concentrations tend t o cause retardation. See Exp. 1, Curve J + (J.

Exp. 1. Action of insulin resp. insulin + glucose on the oxido-reditetion rate in frogs' i,Luacles. Rana ese. 3. The muscles of the hind legs finely minced by cutting; muscle

mass kept on ice.

200 mg muscle mass and 500 y Mb in 1 cc. ~~ K,HPO, in each tube; 3 mol. 100

350 C . in the waterbath.

Fig. 1.

3. ru'either other carbohydrates and carbohydrate relatives (gluco- sides, sugar alcohols etc.) nor substances like lactic acid, the acetone bodies, succinic acid etc. can replace the glucose. While addition of glucose alone (i. e. without insulin), as already mentioned, does not accelerate the oxidation rate, an addition of e. g. fructose, galactose or glycogen causes an acceleration, which is, however, usually reduced by a simultaneous addition of insulin (in a concentration not by itself in- hibiting) - see Exp. 18, 19 and 20. - Only the action of mannose is to a certain extent similar t o that of glucose - see Ahlgren(1) and Bl ix (5).

11*

Page 4: On insulin-like actions of morphine and certain morphine derivatives

156 GLXXAR AHLGREK :

4. The accelerating effect of insulin plus glucose will only appear if the tissue is quite fresh. If the tissue is kept 1.5-2 hours at room tem- perature, no acceleration is noted but a more or less marked retardation may occur (see Exp. 2 and 3? A and B. tube 3). An analysis of this and other phenomenona led t o the development of the theory that a labile agent in the cells, termed gluconzutin, cooperates Ivith insulin and glu- cose in promoting the acceleration [hhlgren (I)] .

I

Tube 11

Ezp. 2 and 3. Insulin cind wzorphine effect inaiaediately nfter the cleccth of the crnincol

and 2 hours late?. Ranne esc. $.

K2H€’0,; 35O C. 3 mol. 200 mg. muscle mass and 700 y mb in 1 cc. __-

100 A. The right hind leg tested immediately after the death of the frog. B. The left hind leg kept a t 190 C for 2 hours, then tested.

I Decoloration time in minutes

36 3 4 . 5

45 I 17 44 1 34

41 40 48 1 26 35.5 34.5 35.5 ~ 25 43.5 , 34.5 36 35

,? i‘ 37.5 37 35 38 37 38 37 37

5. Addition of adrenalin in suitable doses causes an augmentation of the decoloration rate independently of a simultaneous addition of g1ucose.l If both insulin and adrenalin are added in the presence of glucose, an interesting phenomenon of antagonism appears, the tTyo

substances mutually extinguishing each other’s acceleration effect (see Exp. 4, tube 4). This antagonism between insulin and adrenalin is bound up with the presence of glucose, for if no glucose has been added the accelerating effect of adrenalin is undisturbed by the presence of insulin.

When these reactions in vitro are considered in the light of the r81e of insulin in carbohydrate metabolism of the organism as a whole the question arises whether the reactions observed are specific for insulin.

Simultaneous addition of glucose does not seldom impair the accelerating effect of adrenalin.

Page 5: On insulin-like actions of morphine and certain morphine derivatives

03’ INSULIN-LIKE ACTIONS OF MORPHINE, &C. 157

In order to investigate this question I tested the action of about 30 various hormones and drugs, chiefly alkaloids, on tissue oxidation. It appears that the complicated system responsible for the processes of tissue oxidation is very labile. The oxido-reduction rate can be in- fluenced by a great number of substances in oligodynamic concentrations.’ Of the 40 substances tested, only morphine and certain morphine deri- vatives showed an insulin-like effect on tissue oxidation. The other substances which in oligodynamic concentrations influence the oxidation rate in the muscle suspension, differ in their mode of action from insulin in that they act independently of both glucose and “glucomutin”. The substances dealt with in this investigation are as follows.

1. Opium alkaloids. a) belonging t o the group of the phenanthrene derivatiws.

1. Morphine. 2. Codeine (methylmorphine). 3. Thebaine.

4. Papaverine. 5. Narcotine. 6. Narceine.

7. Dionine (ethylmorphine). 8. Heroine (diacetylmorphine). 9. Apomorphine.

b) belonging to the group of the isoquinoline derivatives.

2. Morphine derivatives.

10. Paramorphane (dihydromorphine). 11. Dilaudide (dihydromorphinone). 12. Paracodine (dihydrocodeine). 13. Dicodide (dihydrocodeinone). 14. Monoacetyldihydromorphine.

The substances 1-9 were used in the form of preparations from E. Merck, - Darmstadt; 10-14 were put at my disposal by Knoll & Co.,

Besides the substances which have been tested in experiments published in this paper, the following have also been investigated: thyroxin, hypophysis preparations, cocaine, ecgonine, benzoylecgonine, ergotamine, choline, acetyl- choline, pilocarpine, physostigmine, scopolamine, atropine, ephedrine, histamine, hydrastinine, colchicine, solanine, quinine, coniine, arecholine, aconitine, cheli- donine, nicotine, sparteine, veratrine, digitalis substances, strychnine, brucine, picrotoxine, caffein. Synthalin, the synthetic antidiabetic of F rank , Not h- mann and Wagner does not show the characteristic actions of insulin on tissue oxidation [Ahlgren (lZ)], i. e. it probably is not concerned with the same point of attack as insulin.

Page 6: On insulin-like actions of morphine and certain morphine derivatives

158 GUKKAR AHLGREK :

Ludwigshafeii. The concentrations of the alkaloids in the protocolls are those of the salt used, usually the hydrochloride.

Among the 14 substances mentioned, nio~phin~. corleinP and rlioniw can be grouped together, because, in contrast to the rest, they show the following insulin-like actions :

1. Without the addition of glucose, they produce at certain 101v concentrations a retarding action on the reduction of methylene blue by the minced muscles (morphine: exp. 2, 3, 5-8, 18 and 19; codeine: 18 and 19; dionine: 9, 18 and 19).

2. In thc presence of glucose they produce an accelerating eEfect (morphine: exp. 2-44, 18, 19, 24 and 25; codeine: 4, 18 and 19: dionine: 4, 9, 18 and 19).

3. Fructose and galactose are not able t o replace glucosc (exp. 18 and 19).

4. The action of morphine plus glucose depends, as does that of insulin plus glucose on the activity of the “glucomutin~’. If the “glucomutin” is inactivated, as, for example, by keeping the tissue 2 hours at room temperature, the acceleration is replaced by a more or less marked retardation of the reduction by the addition of morphine with glucose (exp. 2 and 3).

5. These three alkaloids show the same type of antagonism against adrenalin as has been described above for insulin (exp. 4).

Exp. 4. The adrenalin antagonisna.

3m 100

Rana temp. 6. 200 mg muscle mass, 7007 Mb and

Tube 1 ’ Insulin I Morphine ~ Codeine 1 Dionine ]Adrenalin I time in No. I

glucose in 1 cc. K,HPO, ; 350 C. ~- - ___

1 Decoloration ___ -

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17

Page 7: On insulin-like actions of morphine and certain morphine derivatives

159 ON IKSULIN-LIKE ACTIONS OF MORPHINE, kC.

EXP. 5-9. Morphine resp. dionine (without glucose).

Ranae temp. Muscles prepared as in exp. 1.

K,HPO,; 35O (2. 3 mol. 200 mg. muscle maw and 700y Mb in 1 cc. ~ 100

O , I , I I , I . , , I I , , , I

10 - .$

U/ca/oid CORC. - 8

$ -

Fig. 2.

Page 8: On insulin-like actions of morphine and certain morphine derivatives

160 GUXNAR AHLGREN :

The other opium alkaloids act differently. l'hebnitie (exp. 10 and 20), hei,oine (2, 3, 11, 18 and 19), pnpnwerine (12) and apomor.phirie (15)

0 I , , , , I I I , , , I I I I

I0 - s ApomOrpLermc - F

e ,4pomor@me+Fh~mi%

'+ d \ /+-+t,d

1 +/+

'\*-ty /+ 20 -8 Q ,+/+

rm 3u ' ,+ 19eford

40 I ' " ' I '

EX^. 12-14. Paparcrine. Sarcotine. Sarceine.

3 mol. 100

Hanae temp . Muscles prepared as in esp. 1.

K,HPO,; 35O C. 200 nig. muscle mass and 70011 Mb in 1 cc. --

M4 -5 -6 -7 -8 -P -7? a-72 -23 -8 -15 -16 -17 L4 ?V -3,@29

0 1 1 1 , 1 , , 1 1 1 1 1 1 1 1 1

R/c#id conc --

Fig. 5.

Page 9: On insulin-like actions of morphine and certain morphine derivatives

O N INSULIN-LIKE ACTIONS OF MORPHINE, &C. 161

it unchanged or may actually reduce it. Narceine (exp. 14) exeniplifies once more its relative indifference in regard to pharmacodynamic action, only from a rather high concentration (about 10-6) i t causes a moderate retardation, which increases as the concentration rises. Narcotine (exp. 131,

- . .-

i $ 2 w 1 2 3 4 5 6 7 8 9

10 11 12 13 14 15 16 17 18 19 20 21 22 23

- -

E x p . 18 and 19.

Xorph ine derivatives + fructose or galactose. Ranae esculentae. 200 mg. muscle mass and 7007 Mb in 1 cc. 3m

100 K,HPO,; 350 C.

- 1 : 10'2 1 : 1015 1 : 101: 1 : 10'5 -

+2 Q I Decoloration j;- 0 1 time in min. 1 1 3 3 z 8 Exp.lf 5xp.19 w

_ _ _ ~ -

* ' - 35 - 46.5

27 44

- 29

41 - 23.5

47.5

34 43

44

- -

l0/Ll0

l o l o o

-

- -

- j l o loa - , - 26.5 -

lo/oo - - 24.5

43 -

33 lcontrol 39 ' 1 - 26.5 , / + - I 1 -

41 - + 28 + 40 1 - 25.5 + 46 34 0 39 1 - +

-

- - -

25 I - - - 38

-~ +

39 I 28 , + 24 + - - +

*) .+ = acceleration; - = retardation; 0 = no effect.

paramorphane (16) and dilaudide (17) also do not increase the oxidation rate, either by themselves, or together with glucose; above a certain concentration they exhibit a proportional retarding action. Some experi- ments with paracodeine, clicodide and monoacetyldihydromorp~ine (no protocolls published) gave also negative results. Yone of the hydrated morphines showed the insulin-like action.

Page 10: On insulin-like actions of morphine and certain morphine derivatives

162 G 1 r n - s ~ ~ AHLCKEX :

Exp. 20. Thebaine + glucose resp. fructose or grtlactosp.

K,HPO,; 35O C. 3 mol. Runa t e m p . 9. 200 mg. muscle mass and 700 y Mb in 1 cc. - - 100

L'hebaine j Glucose 1 Fructose __ _

Tube n : c

1, 9 & 17 2 & 10 3 & 11 4 & 12 5 & 13 6 & 14 7 & 15 8 & 16

_ _ _ ~ ~ Galactose time in min. + or - 1 Decoloration

- ~ ~ _ _

- 1 : 1013 -

1 : 1013

i : 1013 -

- - I 39; 38; I 0 - - 45; 45; -

- ' 49; 50; - 1 Oleo lo/oo 29; 29.5; I

l o l o o

- - j lo /oo 49.5; 49; -

1O/OO

l - 1 28; 28; + - -

- -

-

11 Nl$$))- 1 Glucose Fructose 1 Galactose

0

20-

u)

w 27 59 90-

1 , / 1 , 1 , 1 , , 1 1 1

Rlca/md conc - + Pominwptwne -

3-3 t ?%G/uco~ * D/oudfde * * +f%oGucooJp

I s

- .E 3 - 3 E -

-$ -

I I , I I I I I I l h f W ,

Paramorp+ Control 4 * 4 * -

_ilRetord,.7hh ~ 4- - _ _ u- -R- -& #Dilaud Cmt$ +

Heroine as well as fructose gave + effect. I n the simultaneous presence of both of them this + effect did not change.

Insulin Morphine Codeine Dionine Heroine Thebaine -- ___

___--_.._I

0 ' + + + +

- ' + - -

- -

- -

01 - - -

Page 11: On insulin-like actions of morphine and certain morphine derivatives

O N INSULIN-LIKE ACTIONS OF MORPHINE, &C. 163

0.1 0.1

Page 12: On insulin-like actions of morphine and certain morphine derivatives

161 Gun-i~ax. AHLGRES :

Microrespirometer experiments. The insulin-like action of morphine observed in the inethylene hliie

experiments was of so great an interest, that it was thought necessary to carry out control experiments with the microrespirometric techniquc. Three series of such experiments were performed, of which details are given below. The tcchnique has been described by Brahnie ( 2 ) and ,\hlgren (3) .

Experiments with Thunberg’s microrespirometers on the action of glucose (0. 2 O i O ) , morphine hydrochloride (1 : lo’), glucose + morphiiir. on the gaseous exchange of 1 gr. minced frogs muscles at 20° C- see p. 163.

One hour after the killing of the animal is necessary for preparation and for establishing gaseous equilibrium. For that reaFon the figures of the gaseous exchange refer only to the second and third hours.

2. The morphine concentration used (1 : 107) is considerably higher than the optimal morphine concentration in the methylene blue experiments (1 : 1012-see exp. 24 and 25). The reason is that in the gas experiment3 the tissue is suspended in the morphine solution only during 5 minutes before the respiration periods, whereas in the methylene blue experiments the tissue is suspended in the morphine solution duiing the ialiolc experiment. (Cf. to Brahme’s discussion of this difference [a].)

3. It might appear at first-sight as if there is a contradiction between the methylene blue experiments, where the acceleration produced by morphine and glucose together does not appear in tissues as old as 1.5 and 2 hours (exp. 2 and 3), and the microrespirometer experiments in which the acceleration effect has been demonstrated during the second and third hours. It should be remembered, however, that in the gas ex- periments the morphine and glucose are added to the tissue about 15 mi- nutes after the death of the frog, that is while the “glucomutin” is still active. The effect of this addition can then be demonstrated in the re- spiration experiment 2 or 3 hours later. As has been shown previously in analogous insulin experiments [Ahlgren (1, p. 168)], the acceleration effect will appear during the third hour also in methylene blue experiments if only the insulin and glucose are added together to the tissue in an earlier stage, before the glucomutin gets inactivated. Accordingly, there is no contradiction a t all between the two kinds of experiments.

4. Four microrespirometers were used, and they were employed in turn for control, morphine, glucose, and morphine + glucose experiments (see the table). For that reason the strong increase in the gaseous ex- change after addition of morphine plus glucose cannot be attributed to a constant error in one of the instruments.

As the table shows, the results of the microrespirometer experiments fully confirm the results obtained with the methylene blue method. Whereas in all the experiments the gaseons exchange ot the control,

Coini/wzt 1.

Page 13: On insulin-like actions of morphine and certain morphine derivatives

O N INSULIN-LIKE ACTIONS OF MORPHIWI’:, kc. 16 5

morphine and glucose experiment shows practically the same ralucs, the oxygen consumption, as well as the carbondioxide production, is strongly augmented in the tissue preparation which had been treated with morphine plus glucose. This augmentation is connected pith an obvious increase of the respiratory quotient, but the significance of the increase is not clear. In tissue respiration experiments there is an even greater need for caution when attempting t o interpret changes in the respiratory quotient than in experiments on intact organisms. I refrain from discussing the point in question in this paper.

The relation between the chemical constitution and the insulin-like action of the morphines.

It is firstly of interest to notice that the insulin-like action has becn observed only in substances belonging t o the group of the phenanthrene derivatives. The three opium alkaloids of the isoquinoline group did not show the property in question.

Fig. 7 represents the constitution formulas of some of the phen- anthrene derivatives tested. The nmphine formula (a) is represented according t o Schopf (13).

In the morphine formula there are 3 0-atoms: the first (very indifferent) one bound t o the carbon atoms 4 and 5, a second one contained in the phenolic OH-group at 3, and a third one in the alcoholic >C<& group a t 6.

Codeine and clionine differ from morphine only in that \Tag that the phenolic OH-group has been niethylated in codeine (b); ethylated in dionine (c). These substitutions have not deprived the molecule of its insulin-like action on the tissue oxidation.

Heroine is diacetyl morphine, both the phenolic and the alcoholic OH- groups having been acetylated. This change causes a disappearance of the insulin-like action. The case of thebaine (e), in which both the OH- groups have been methylated, and two hydrogen atoms have been re- moved, is similar. It is therefore easily understood that the insulin-like action disappears when those radical changes take place which charac- terize the transformation of morphine into apomorphine ( f : according to P s c h o rr).

Puramorphane (9) is dihydromorphine, the double bond between carbon atoms 7 and 8 having been reduced. In the same way para- codeine has been derived from codeine. Through this reduction of the morphine or codeine the insulin-like action is lost so that it is not sur-

Page 14: On insulin-like actions of morphine and certain morphine derivatives

166 GUNKAR XHLGREN :

a) Morphine (ace. to S e h o p f , 19273

H a) Heroine

H

Y g) Paramorphane

b) Codeine c) Dionine

H

H O

HO

H N.CH, I

CHZ H

H

(am. to 8 chopfj e) Thebaine f) Apomorphine

(ace. to Psehorr)

H

Fig. 7.

N CH, I

o@H 0. 6 HZ C H 2

h) Dilnudide

Page 15: On insulin-like actions of morphine and certain morphine derivatives

O N INSULIN-LIKE ACTIONS OF MORPHINE, k C . 167

prising to find that this action is also not shown by nzonoacetyldihydro- morphine, dilaudide (h ; dihydromorphinone), and aicoaide (dihydro- codeinone). In the two last substances, besides the reduced grouping, the secondary alcohol grouping has been exchanged for the carbonyl group.

It is clear, therefore, that the examination of the relation between the insulin-like action of the morphine derivatives and their chemical constitution has so far shown that the insulin-like action i s associated with the presence of

1. the >C<iH -group at the carbon atom 6 ; 2. the double bond bettceen the carbon atonzs 7 and 8. 0% the other hand a substi tution of methyl or ethyl in the phenol group

It is proposed to extend the investigation t o other morphine and (a t3 ) does no t Prcludr u n insulin-like action.

phenanthrene derivatives when occasion offers.

On morphine and opium a8 antidiabetic agents. The insulin-like action of morphine observed in the in vitro-experi-

ments described gains in interest when it is considered in the light of the therapeutic use of opium preparations in diabetes mellitus.

For centuries [see K a u f m a n n (14)] the use of opium has been re- commended in this disease, and, in deed until the introduction of insulin the drug stood first on the list of antidiabetics. Naunyn wrote in his diabetes monograph 1906 (15, p. 433): :,Die Zahl der empfohlenen Mittel ist Legion, und fur ihre Mehrzahl fehlt es nicht an ,Belegen', d. h. man sah unter Gebrauch der betreffenden Droge die Zuckerausscheidung her- untergehen. Doch gibt es meiner Kenntnis der Dinge nach kein einziges Medikament aul3er dem Opium, welches einigermal3en zuverlassig eine giinstige Wirkung auf die Glykosurie ausubt." Also in 1917 v. Noorden (16, p. 374) stated : ,,Den . . . immer wiederkehrenden Angaben, da13 Opium die Zuckerausscheidung vermindere, kann ich mich mit zahl- reichen Beweisstucken anschlieflen." A particularly good effect on the acidosis was observed by Landergren (17) and confirmed by v. Noor- d e n (16). Reviews on the literatur in this field are found in the papers of K a u f m a n n (14) and af Klercker (18). Even as late as 1921 a diabetes clinician so experienced as P e t r e n (19) used opium medication (7-10 drops tinctura opii 3 times a day) in all severe and medium-severe cases-a statement which Jos l in (20, p. 529), however, when relating it, apostrophizes with an ( ! ) !-With the beginning of the employment . of insulin opium completely lost its importance as antidiabetic; in Ber- tram's review ,,uber die medikamentose Behandlung des Diabetes mel- litus" (21) 1928 opium ist not even mentioned.

Page 16: On insulin-like actions of morphine and certain morphine derivatives

168 GUNNAR AHLGREN :

For several years attcnipts have been made to explain the anti- diabetic effect of opium as resulting from the delayed absorption of food in the intestine, caused by the morphine Contraction of pylorus, although also other hypotheses have been discussed [see e.g. Y a u n y n (15)l. 1915 af Klercker ( l8) gave experiniental evidence in favour of the “delayed absorption theory”, but at the same time he had t o conclude from his analysis of blood sugar curves in diabetes patients after opium supply that o p i z m nlso seems to exercise (I, specific cict ion ngninst the d in- betic nzetnbolisni disturbance.

Experiments by Cammidge, F o r s y t h and Howard (23, 1922) on the influence of morphine on the hyperglycemia in dogs and men gave no information as to the mode of action. In 1923 Holm (22) published the results of experiments on dogs and men showing that the delayed absorption plays an important r81e in the depressing action of morphine on alimentary hyperglycemia, but they do not exclude the other possible actions.

The question of the antidiabetic action of opium and morphine has been very much complicated, however, through the observation that opium (as well as morphine) in many cases bring about not a de- crease of the blood sugar but an increase. This does not necessarily exclude the possibility that the substances in question arc able to exert an insulin-like, antidiabetic action, for it must be recognized that the complex morphine molecule attacks the organism at more than one point.

Some authors [Cannon, Mc I v e r and Bliss, also Abe-see Danielson (25); S t e w a r t and Rogoff (%)I have tried to relate the morphine hyperglycemia to a stimulation effect on the suprarenal medulla, which considerably increases its adrenalin secretion under the influence of m0rphine.l After epinephrectomy (on cats) S t e w a r t and Rogoff (23) found that morphine injection caused little or no hyperglycemia. In experiments on epinephrectomized dogs Holm (22) observed a slight increase of the blood sugar after morphine (or a t least retarded fall of the blood sugar curve). He inclines to locate the attack of morphine in the liver or the liver nerves.-Marenzi (28) reported in 1926 that splanchnec- tomy considerably reduces the morphine hyperglycemia in dogs. After complete abdominal sympatectomy he noted that morphine injections produced a fall of blood sugar instead of hyperglycemia.

Danielson (25) has studied the effect of a simultaneous injection of insulin and opium (pantopon) on the blood sugar of rabbits. He found that a large dose (50 mg. pro kg.) weakened the lowering effect on the blood sugar of a certain insulin dose (1 internat. unit pro kg.). Smaller doses

Several authors ascribe to insulin itself the ability of stimulating - directly or indirectly - the suprarenals to increased adrenalin secretion. See e. g. Choay’s insulin monograph (24, p. 449).

~ ~~

Page 17: On insulin-like actions of morphine and certain morphine derivatives

ON INSULIN-LIKE ACTIONS OF MORPHIXE, &C. 169

(0.5 to 7 . 4 mg. pro kg.), which by themselves slightly increased the blood sugar, did not diminish, but seemed sometimes to increase, the hypo- glycemia effect of the insulin.

Perhaps it should be mentioned in this connection that Arne th (29) has reported in human diabetes, ,,da13 die Insulinwirkung dann durch die (langst bekannte) die Entzuckerung begunstigende Opiumwirkung un- verhaltnismafiig stark unterstutzt wird".

It is obvious, however, that further investigations are needed in order to clear up the question of opium as antidiabetic; as for example, the study of the gaseous exchange or acidosis as indicators of the anti- diabetic action of opium and morphine [see Ewer t and Ahlgren (30)].

The question as to which of the constituents of opium are responsible for its antidiabetic action has also been discussed. Clinicians have long ago tried the various opium alcaloids in this respect. In view of the findings reported in this paper it is of interest that only morphine and codeine have been recommended as antidiabetics ; indeed some French investigators have preferred them to opium. Papaverine and narcotine were cancelled [cf. Kaufmann ' s compilation (14)].

An attempt to determine quantitatively the insulin-like action of morphine.

Theoretically the insulin-like action of a certain quantity of mor- phine ought to be measured in insulin units. From the preceding account it is evident that a determination on the basis of the blood sugar is im- possible. An attempt has, however, been made in the following manner: The influence of a series of various insulin or morphine concentrations on the oxido-reduction rate in minced muscles from one and the same frog in the presence of lo/,,,, glucose (exp. 24 and 25) has been investigated. The curves in fig. 8 and 9 illustrate the results.

In both the experiments the maximal acceleration effect of insulin (the shortening of the decoloration time from 62 t o 37 min. or from 41.5 t o 24 min.) has been somewhat greater than that of morphine (from 62 t o 41 min. resp. from 41.5 t o 27 min.). More important is, however, that the optimal insulin concentration (1 : IOl3 in both the experiments) is about 100 times lower than the optimal morphine concentration (1: loll); thus, the insulin preparation used is more active than mor- phine hydrochloride. If one considers the distance between the insulin and the morphine curves in whole their length (they run practically parallel), a very tentative estimate might be advanced that the insulin preparation used were 10-100 times as active as morphine hydrochloride, that is 1 mg. insulin (= 15 internat. units) = 10 t o 100 mg. morphine hydrochloride. This estimate is naturally made with due reservations.

Skandinav. Archiv. LVIII. 12

Page 18: On insulin-like actions of morphine and certain morphine derivatives

170 G~NNAR AHLGREX :

EX^. 26-25. Qztantitatice comparison between the insulin and the tnorphine ej'ecf.

Ranae temp. Muscles prepared as in exp. 1.

200 mg. muscle mass, 6007 Nb and lo/ofl glucose in 1 cc. 3 mol.

100 - K,HPO, in each tube: 350 C in the waterbath.

6

fa

20

3

40

59

do

70

80

Fig. 8.

Fig. 9.

Page 19: On insulin-like actions of morphine and certain morphine derivatives

O N INSULIN-LIKE ACTIONS OF MORPHIEE, c ~ C . 171

On this basis it is possible to estimate the order of magnitude of the antidiabetic effect of the opium doses used in the diabetes therapy; it must be recognized that the fact that my experiments were made on frogs muscles will render the estimation even more uncertain. 8 drops of tincturu opii x 3 = 24 drops pro die = 0.6-1.0 gr. If we estimate as lo/, the content of active constituents (morphine and codeine) the dose pro die will correspond to about 6-10 mg. morphine.

On the basis that insulin is 100 times more active than morphine, the daily dose of opium tincture would correspond to 0.9-1.5 international insulin units. If the insulin is only 10 times more active, the same daily dose would correspond to 9-15 units; R reasonable antidiabetic agent.

The results of this investigation raise naturally two important questions.

In the first place, is it possible t o produce a phenanthrene deri- vative which possesses the insulin-like action of the morphine, but which is without its narcotic action ? And secondly, is there a chemical relation- ship between morphine and insulin?

Summary.

1. The author has previously shown that insulin performs a series of typical reactions in experiments on tissue oxidation (methylene blue method; microrespirometers), reactions which with all probalility are intimately connected with the primary physiological function of insulin.

2. When about 40 various substances were tested as to their ability to replace insulin in these reactions, it was found that only morphine and certain morphine derivatives (codeine and dionine) showed an insulin-like action. Other opium alkaloids (thebaine, papaverine, narco- tine, narceine) and morphine derivatives (heroine, apomorphine, dihydro- morphine, dihydromorphinone a. 0.) did not show the reactions in question.

3. The relation between the chemical structure and the insulin- like action of the morphines is discussed.

4. The insulin and morphine effects on tissue oxidation have been compared quantitatively.

5. The observation that morphine can exert an insulin-like action gains in interest in light of the fact that opium has for centuries been used in the treatment of diabetes. The question of the antidiabetic action of opium and morphine is discussed.

_ _ _ _ ~

This investigation was aided by a grant from “Stiftelsen Therese och Joh a n An der s s o ns Minne”.

12 *

Page 20: On insulin-like actions of morphine and certain morphine derivatives

172 GUSSAR AHLGRES: 0s ISSULIX-LIKE ACTIOSS OF JIORPHISE. C ~ C .

Literature. 1. 0. Ahlgren, Thi s drchiv. 1925. Bd. XLTII (Suppl., p. 1-266). 2. L. Brahme, Acta nzed. scand. 1925. Vol. LXII. p. 188. 3. G. Ahlgren, This Archiz.. 1926. Bd. XLVII. p. 271. 4. J. J. R. Nacleod, “Carbohydrate .Mefubolisnr and Insttlix”. London

5. G. Blix, Thi s Arckiv. 1926. Bd. XLVII. 1). 292. 6. A. Gigon, Ergebn. d . Physiol. 1925. Bd. XXIT’. p. 196. 7. S. A. Holboll, This Archiv. 1926. Bd. XLVIII. p. 225. 8. U. Y. Euler , Pflugers Arch. f . d. ges. Physiol. 1927. Bd. CCXVII.

9. €1. J. Wolf, Ibidem. 1927. Bd. CCXVI. p. 322.

1926. p. 321.

p. 699.

10. G. Ahlgren, Abderhalden’s Handbitch d. biolog. Arbeifsmefic. 1926.

11. T. Thunberg , Oppenheimer’s ,,Die Fermente und ihre Wirkungen“.

12. G. Ahlgren, Biochem. Zeitschr. 1928. Bd. CCVI. p. 99. 13. C. Schopf, Liebig’s Ann. d . Chewa. 192i. Bd. CDLII. S. 211. 14. M. Kaufmann, Zeitschr. f. klin. Med. 1903. Bd. XLVIII. S. 260. 15. B. Piaunyn, ,,Der Liabetes naellitus“. 1906. 2. Aufl. 16. C. I-. Noorden, ,,Die Zuckerkrankheit und ihTe Behandlung“. i. Aufl.

17. E. Landergren, XOrd. M e d . Arkiv. Afd. 11. 1910. Vol. XLIII.

18. Kj. 0. af Klercker, Deutsch. Arch. f . klin. Ned. 3915. Bd. CXVIII.

19. K. PetrBn, ,,Diabetesstudier“. Kopenhamn 1923. 20. E. P. Joslin, “The treatment of diabetes mellitus”. 3rd ed. Phila-

21. F. Ber t ram, Klin. Tl’ochenschr. 1928. 7. Jg. S. 1209. 22. H. Holm, Zeitschr. f. d . ges. ezp. Med. 1923. Bd. XXXVII. S. 81. 23. P. J. Cammidge, J. 9. Cairus For sy th and H. A. H. Howard,

Lancet. 1922. Vol. C. 2. p. 1324. 24. A. Choay, ,,La secretion interne de pancreas et l’insuline“. Paris 1926. 25. C. G. Danielson, Compt. rend. d . s. d . 1. SOC. d. biol. 1926. T. Vc.

26. G. N. S tewar t and M. Rogoff, Journ. of pharm. a. exy. ther. 1922.

27. The same, Amer. Journ. of Physiol. 1922. Vol. LXII. p. 93. 28. A. Marenz , Compt. rend. d . s. d. 1. Soc. d. biol. 1926. T. VC. p. 1171. 29. Arneth, Klin. Wochenschr. 1925. 4. Jg., S. 1169. 30. B. Ewer t and G. Ahlgren, I n manuscript,

Abt. IV: 1. S. 671.

1929. 5. Aufl. Bd. 111. S. 1118.

Berlin 1917.

Hafte4. p. 1.

S. 85.

delphia 1923.

p. 1058.

Vol. XIX. p. 59.