US2154432A - Therapeutic colloid - Google Patents

Description

Patented Apr. 18 1939 PATENT OFFICE THERAPEUTIC COLLOID Garth Wilkinson Boericke and William Wallace Young, Philadelphia, Pa., assignors to J. C. Shay, Incorporated, Philadelphia, Pa.

No Drawing. Application April 27, 1936, Serial No. 76,602

4 Claims.

Our invention relates to the art of the production of substances in colloidal state to be used therapeutically and more particularly to the production of fatty sols that adsorb the toxins that 5 pervade the human blood stream when the body is in a state of unhealth.

The object of our invention is the preparation of stable fatty sols by a novel process which can be administered intravenously or subcutaneously 10 for the purpose of adsorbing and fixing the toxins that are frequently the evidentiary manifestation of disease and for the ultimate purpose of restoring the health of an individual to an accepted.

- I which the name Blooddust had been generally applied. One of the early observers had previously suggested the term Chylomicron for these fine particles, inasmuch as investigation had established their fatty character and approximate Jo size, one-half to one micron; other facts established by the early workers were that chylomicrons had their origin from fat and not from protein or carbohydrates, that the number circulating on the blood stream has a constant relaj tion tothe food intake and the approximate nu- V merical relationship to other formed elements were estimated, i. e. in a cubic millimeter of blood there are about eighty million chylomicrons to five thousand White and five million red. The m last point seems to us specially important,the tremendous'quantity of these particles in relation to the blood cells. It is obvious that such a predominance of one subject in the blood serum should have more biological significance than 5 simply the function of nutrition and hence might logically be affected by the various states of health and disease in the same manner that the red or white cells are. 1

Search of the literature for further .of this important emulsion brought to light a series of papers by certain workers whose careful work concerned itself with the physical and chemical aspects of the 'chylomicron emulsion. They concluded that each fatty particle was ll coated with-a protein film, presumably a mixture knowledge ol albumen and globulin and further that the number of the circulating particles had no effect on the specific gravity or the viscosity of the blood as shown by the methods they employed in their work.

Further research revealed that the extension ultramicroscopic investigation of the blood serum has been done by certain other individuals. These workers consider the electrophysical state of the particles to be the determining factor in health and disease and utilizing the modern conception of the structure of the atom they feel that an explanation of all diseased processes is possible by their hypothesis. The essential fatty character of these minute particles is not recognized by these last mentioned individuals but then, as has been shown previously, the surface film may well be protein, this being a major factor in the stabilization of such an emulsion.

From this work, there is no doubt that there is a very suggestive relationship between the state of health of the individual and the morphology and characteristics of the small particles.

We have examined over a thousand cases under the dark field covering a wide range of sickness and it is quite possible to describe an illness in terms familiar to the physical chemist by observing the ultra-microscopic picture of a blood serum.

Before discussing the common changes observed under the dark field, a few definitions should be in order:

Brownian movement-This is a vibratory motion peculiar to small particles in a state of colloidal suspension. It is due apparently to the electronic bombardment of the particles by the external phase.

Hydration.-Where the field shows a number of variations under different heads, for examplea fewer number of particles-an increased size of the particle-a sluggish repulsion between particlesa sluggish Brownian movement-the presence of clumps.

Dispersion is the opposite of the above and is identified generally with the normal appearing field-that is a goodly number of particles, excellent motion, usually uniform in size, and no clumps.

After stating the possible biological variations in the characteristics of a fatty emulsion, we must note some of the more obvious changes, commonly observed by looking under the dark field. The commonest findings are a lessened activity and diminution in the number of these circulating chylomicrons, especially in wasting diseases. In diseases of an endotoxic character, clumping and agglutination under the miscroscope of these individual particles seem to be a sign of infection or of existing toxemia. This clumping and agglutination by the fat particles in the presence of a circulating toxin suggests that perhaps fat plays a part in detoxification. In other words, fats are toxiphylic and fix a toxin, the union being chemical or physical, and resultinginan innocuous substances.

The ability of a lipoid and fat to fix toxic radicals is well known, and certainly the colloidal form circulating in the blood is, by virtue of its tremendous surface development, singularly well adapted to this role.

Some examples of this power of fat toabsorb may be given: the aflinity of diphtheria and tetanus for the lipoids of our body. If wedelay in giving antitoxin for instance, there will be no unbound toxins circulating to meet it and, hence, the danger of delay in giving antitoxin is serious.

Again, We know that a prolonged toxemia of an organ leaves a deposition of fat in it. Fatty degeneration and infiltration are every day experiences to the pathologist. A kidney which has lost its function through inflammation is apt to'be full of fat, a liver likewise reacts by having fatty replacements, especially in severe poisoning. It was formerly believed that this depended on a'transformation of-the'proteins of 'the tissues into fat, under the influence of the poison. It is evident that if such a change were as wide-spread asthe fat deposits would lead chematous cells is not formed from their own protein, but is deposited after transportation from fat deposits already existing.

"Our point about this whole matter is-the assumption that a toxic organ has 'fat brought to'it by'nature in an effort to detoxicate the existing condition and is not'simply a phonon enon'per se of the action of toxin on cells.

It'may not be an exogenous type like diphtheria or tetanus. It may be endogenous'orintracelldlanexerting its eifects only upon cell destruction. 'It may be on the other hand an untoward byeproduct of deranged metabolism, or finally and most probably, it may be a so-called change of physical or chemical equilibrium and not'a separate substance at all.

We have prepared lecithin sols but consider lanolin superior, which is a tri-cholesterol-ester of'the three fatty acids palmitic, stearic and oleic. 'It is produced by condensation methodsusing alcohol and water, and when finished is extraordinarily stable, but extremely sensitive to a toxic radical. The actual method of procedure is indicated as follows:

Anhydrous lanolin is used because itis slightly soluble in alcohol and partly mixes with water without separating.

Quantitative measures are not necessary in the preparation of this sol because there is only a certain amount absorbed by the alcohol, the rapidity and quantity varyin with temperature of'the alcohol. For this reason warm alcohol is used, to save time and conserve alcohol, though itmay be made with cold. A. sufficient amount,

movements. 'ingthe gen'eralrule that the phase'having the lower dielectric constant carries a negative charge, the positive phase in this case being group turned toward the water.

usually about fifty grams of lanolin is stirred in a large beaker by means of a mechanical stirrer for one and a half to two hours. Samples are taken after the first hour and mixed with an equal quantity of distilled water. If a prompt, white precipitate forms, which alter several samplesdoes not increase in density, this is taken as evidenced sufficient saturation.

The volume of lanolin-alcohol mixture is then mixed with three times its quantity of distilled water and an immediate white emulsion is formed.

The next step is the driving ofi of the alcohol over a steam bath at a temperature varying from eighty to ninety degrees. The volume at the end of this process is roughly half that of the original 'mixture and alcohol should not be detected by smell although there is a small quan- 'l'iave shown it to be sterile from all types of organism and'spore formers. Freezing promptly breaks the emulsion, and agitation fails to restore it.

It may be stored in a cool place with acork 3 or cotton plug. Before clinical use, it is brought to a boil or is autoclaved,-and injected slowly at body temperature.

Its'physical properties are: a milky-appearing aqueous'liquid, consisting of particles about one micron in size showing active Brownian It carries a negative charge obeywater. its pH varies slightly, but may be adjusted easily to 5.5. There is undoubtedly an orientation of the fatty molecules'with the polar Hence, any toxin with which it united must present a similar active or polar group as recognized by colloid chemistry.

We contend that our emulsion is not stabilized by any film, dispersion being quite permanent even after three years observation. its stability depends upon two factors: one being-the electric charge and the other a hydration factor-spoken of by Freundlich.

The sol is thermo-stable even at boiling temperature and ultra-violet light rather increases the energy of the Brownian movement.

In a general way, dispersion is enhanced "with an increase of thepH or a shift to the basic side of the solution, While hydration is brought about by the addition of acids. 100 cc. of colloid by 30 minimum lethal doses of diphtheria toxin'will occur inlanolin sol at a pH value of 4.5, but will not occur atapI-I value of 5.0. Most diseases of a toxic nature are associated generally with acidosis which is compensated for by the buffer salts. In. these diseases,.we note routinely a state of marked hydration of the chylomicrons in the blood serum. Conversely in diseases characterized by alkalosis,fthe chylomicrons in the serum are routinely highly dispersed.

Presumably,

Thus,.precipitation of The effect of a number of substances upon this colloid was investigated next. From test-tube experiments, we demonstrate that: a drop of toxic blood will precipitateapproximatelya cc. of this lanolin. Toxic blood is meant simply that the blood comes-from a case showing ordinary-symptoms of toxemia, such as fever, leucocytosis, malaise, sweat, etc.

Normal blood will notdo this. None of thefour blood typestaken from-a well person has any precipitating effect on the colloid. We have tried because it is the presumed afiinity of the fat for.-

toxin which causes a precipitation; and ifmany other substances did this, it would be of no par ticular value. Other common chemical substances do not do this. Thus methylene blue (10%) and phenol (97%), arsenic (1-100), mercurochrome (2%), bichloride ofmercury (10%) S. T, 37, ceanothyn (tincture), iodine (sat. sol.), sodium hydroxide (16%) have no precipitating effect according to our experiments.

Other observations follow: Precipitation is caused regularly by a pH'more acid than 1.2; but acidity is not the only factor as we tested many sera which were definitely on the alkaline side, and a drop-of which promptly precipitated the colloid. The promptness with which coagulation or precipitation takes place in the test tube after the addition of a drop of toxic bloodfrom a sick patient is roughly parallel to the severity of the illness. Thus in a critcial pneumonia case in the first stage, precipitation was prompt in 4 minutes and when the test was repeated 1 week later, with the patient considerably bettenthe test took 20 minutes before precipitation was noticeable. This phenomenon observed in vitro may be interpreted much as the blood sedimentation testfor red cells. 7

Of the various salts, the Cl ion seems to be particular in causing precipitation in a lower concentration than other salts- Regarding this precipitation or agglutination phenomenon we assume that the fat fixes a toxic radical; and inour test-tube experiments this precipitating effect is considered evidence of toxicity. When acid is the precipitating agent the result is due to'the H charge neutralizing the negative charge on the particles.

Analyzing this phenomenon, it is clear that if the toxin in a toxic blood hasnot'been fixed by the emulsion in thetest-tube, the precipitate "if used again against another specimen ofcolloid in a test tube would drop it. But this will We found first that the colloid in 5 to 10 cc. doses harmful to humans.

was not harmful when injected into the earveins of a rabbit and doses of 250 cc. are not The next step, we felt, was then to obtain a definite toxin. of known strength instead of simply an unknown toxic serum. We

' obtained from a local laboratory diphtheria toxin represented this dose.

otherseries of experiments, the conclusionsyof which follow:

Thefatal dosewhenmixed with-3 cc. of colloid andinjected together was without efiect. The control animal diedon thefourth day.

Three guinea pigs were given a lethal dose of" toxinv subcutaneously and then, each was given respectively 1, 2, 3, cc. of colloid in divided-doses starting 15 minutes after the original toxin injection and continuing every 2 hours for, 4 days. The results were that the guinea pigs treated. bythe-colloids showed noeffects so far as We couldjudge by their appearance and behavior. The control animal died as before.

Further experiments brought out the fact that it takes more colloid to neutralize the toxin in the body.=than in the test-tube and also that if, given all in one dose it is not as effective as the same amount divided over a period.

Totest out the detoxicating power of fat in vivo, 3- minimum lethal doses of toxin were put into 10cc. of colloid and allowed to precipitate. At the end of 24 hours, the clear portion was injected into av guinea pig and the precipitated portion was injected into another guinea pig. In both pigs no untoward effects were noted, which would'seem to indicate that the toxin had been adsorbedby the precipitate rendering both clear portions and precipitate innocuous. Naturally, many experiments have been made with the end in view of shaking loose the toxic portion after its union with the fat, all being unsuccessful to date.

This however opensup a most fascinating investigation because, were it accomplished, one would be in a position of being able to isolate an endogenous toxin, such as that of pneumonia, typhoid; grippe, etc., and. study it in its pure state as we do now'diphtheria, tetanus, and botulinus.

In the preparation of this colloid for human injection, it was necessary to find out whether or not the buffer salts have a precipitating effect.

The addition'of phosphates, sulphates and calciumcarbonate to the lanolin sol does not influence the state of dispersion, the Brownian motion, the-sizeof the particles, or its therapeutic efficacy in any way until a certain concentration is reached. Since this concentration is well above that found physiologically, it is possible to employ a sol to which has been added one or more of these salts in physiological proportions. The addition of sugar in the form of dextrose up to a l0'per cent solution with or without the presence of a salt, such as sodium carbonate, in no waymilitates against the stability of the sol. This finding is of value since it permits the use of a physiologic glucose solution (5 per cent).

Unadjusted, when freshly prepared, a colloidal sol of fat will be of a certain pH value. For instance, lanolin will have a pH value of about 4.5 or a little higher. This is not the pH of the body, but from our experience gives excellent therapeutic results and even when used in as much as a 500 cc. dose has no deleterious effects. It is wise, however, and it is our custom to adjust our sols by the use of small quantities of a very dilute sodium hydroxide to: a pH of 7.5 or 7.6 before we use them. It has been found that, if a sol of lanolin has a pH of 4.5, the addition of sugar to make a 5 per cent solution will raise the pH value to almost a physiological normal due to apparent depression of ionization. For this reason, it is hardly necessary to adjusta sol to which glucose has been added. Somewhat the same effect, although not to the same degree, follows the introduction of salt or sodium phosphate in a physiological percentage.

Our sols have routinely the appearance of milk and have a surface tension slightly'above that of water. In each 1000 cc. there is approximately 4.5 to 5.0 grams of fat. It is practically odorless and produces nausea when an attempt is made to dring it.

Therapeutically, there is a choice among a number of preparations of fat. We have used lanolin almost exclusively, although kekuni nut sol is just as efiicacious. However, almost any other fat can be used. The sol can be used in its freshly-prepared state or with the addition of dextrose. The method of administration depends upon the preparation to be used. With the freshly-prepared sol, hypodermoclysis is the method of choice and as much as 250 cc. will be absorbed readily in minutes. If dextrose or the physiological salts or both are to be used, then the intravenous method is to be preferred, since the hypodermic method leads todiscomfort because of the slow absorption. The intravenous use of the freshly-prepared specimen will at times cause severe lumbar pains after some to 50 cc. have delivered. Since the material is kept as described, it is wise to boil the sol for 3 minutes before using, and to administer it at body temperature.

We have noted no sign nor symptoms of an allergic nature even of the most minute character in cases that we have treated. The initial dose is from 100 to 300 cc. depending upon the degree of toxicity. The subsequent doses, if necessary, are given at 2 to 5 day. intervals, depending upon the progress of the case. Where there is marked dehydration, anorexia, or imperfect nutrition, we find it wise to use the sol containing sugar. Where the toxemic manifestations alone predominate, we merely use the adjusted colloid. The indications for its use are the infectious diseased, from the mild cases exhibiting a low-grade toxicity which show merely a generalized aching and soreness and tend to become chronic to the most virulent type of infections with temperatures ranging as high as 107- degrees. Among the cases treated successfully the streptococcic and staphalicicin infections, cellulitis, septic arthritis, puerperal sepsis. Also we have observed the toxic symptoms of tuberculosis improve after colloid injection (usually sub-cutaneous).

Clinical toxic symptoms of the case serve as an indication, rather than as a definite diagnosis. Also, since the affinity of the fat for the toxin is not bacteriologically specific, the type of infecting organism is not of very great importance, and the colloid may be indicated equally well in a case of typhoid, as in a caseof pneumonia or meningitis. The best and most scientific indication for its use is to be had by a study of the serum under the dark field; and if hydration and clumps are marked, it is indicated.

Following the use of the lanolin with dextrose intravenously, there is frequently met with a febrile reaction with a rise of temperature of 1 or 2 degrees and a concomitant increase of the pulse rate. This is followed usually by a decrease of the temperature to normal or thereabouts. There may be in 24 or 48 hours another increase of temperature, but it is wise to wait for at least '72 hours in a subacute case, longer in a chronic case, or 24 hours in an acute case. Before repeating the dose, the indications for repetition are found in the behavior of the temperature curve. The febrile reaction from a dose of 200 cc. intravenously is marked by chills, fever, and sweat which last from 15 to 20 minutes. This does not occur in all cases, however, and the febrile reaction is avoided by the hypodermoclysis method of administration. Contra-indications have as yet not been met with, although the use of the colloid is apparently contra-indicated in athermatous subjects and in those under heavy digitalis dosage.

We are convinced that the colloid has no effect directly on bacteria themselves, acting simply as a therapeutic sponge to absorb cellular and bacterial toxins and poisons and thereby give the patient a chance to capitalize on his natural resistance or the other treatment instituted.

We are aware that the invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof, and we therefore desire the present embodiment to be considered in all respects as illustrative and nonrestrictive, reference being had to the appended claims rather than to the foregoing description to indicate the scope of our invention.

We claim:

1. A process for preparing lanolin sol which consists in agitating lanolin with ethyl alcohol, mixing said lanolin and said ethyl alcohol with approximately three times their volume of distilled water, expelling substantially all of said ethyl alcohol by heat, and filtering the hydroalcoholic emulsion of said lanolin.

2. A detoxicant, therapeutic compound for parenteral use comprising lanolin sol of colloidal particles of substantially 1 micron in size and unstabilized by other substances.

3. A detoxicant, therapeutic compound for subcutaneous use comprising lanolin sol of colloidal particles of substantially 1 micron in size and unstabilized by other substances.

4. A detoxicant, therapeutic compound for intravenous administration comprising glucose and lanolin sol of colloidal particles of substantially 1 micron in size and unstabilized by other substances.

GARTH WILKINSON BOERICKE. WILLIAM WALLACE YOUNG.