WO2001010252A1

WO2001010252A1 - Anti-oxidant inhalant and method

Info

Publication number: WO2001010252A1
Application number: PCT/US2000/021902
Authority: WO
Inventors: Robert P. Lenk
Original assignee: Therapeutics 2000, Inc.
Priority date: 1999-08-11
Filing date: 2000-08-10
Publication date: 2001-02-15
Also published as: CA2391728A1; EP1239745A1

Abstract

This invention described here is designed for ameliorative or prophylactic use by smokers or others who are at high risk of developing environmental induced lung disease. In one embodiment this involves a cigarette filter-type device (24) comprising a pharmaceutically acceptable high-effect antioxidant such as BHT or BHA provided in an inhalable therapeutically effective amount, and a method of manufacture and use.

Description

ANTI-OXIDANT INHALANT AND METHOD

Field of the Invention

The invention described here is designed for ameliorative or

prophylactic use by smokers or others who are at high risk of developing

environmental induced lung disease. In one embodiment this involves a

cigarette filter-type device comprising a pharmaceutically acceptable high-

effect antioxidant such as BHT, propyl gallate and BHA provided in an

inhalable therapeutically effective amount, and a method of manufacture

and use.

Background of the Invention

Damage caused by environmental aerosols, tobacco and other

smoking products is clearly established in the literature. Chronic smoking is

associated with a number of health problems including cardiovascular

disease, lung cancer, and local damage to the lung tissue which eventually

develops into chronic obstructive lung disease. The term chronic

obstructive lung disease is to be broadly understood to encompass

emphysema, pulmonary fibrosis and a number of other chronic lung

diseases.

As many as 1 77,000 Americans were diagnosed with lung cancer in

1 998. Of those, about 1 06,000 were men and 71 ,000 were women.

Unfortunately, lung cancer usually remains undiscovered until a tumor has grown large enough to compromise respiration or another physiological

system, at which point the prognosis is poor. Experience indicates that

only 1 out of 8 of patients diagnosed with primary lung cancer survives 5

years.

Resources have been expended on developing new treatments for

lung cancer. Abstinence counseling has been ineffective in reducing

smoking among teenagers, and teenage smoking has been found to lead to

lifelong addiction. Little effort has been directed toward preventing lung

cancer in continuing smokers. The reality is that, despite the active

education campaign, 48,400,000 adults and 4, 1 00,000 teenagers in the

United States continue to smoke (1 .1 billion smokers worldwide). In 1 996,

487 billion cigarettes were consumed in the US.

Without being bound by any particular theory, it is believed that a

malignancy results from a multi-step process; a cascade of events. Some

researchers believe that tumors evolve from a single cell that has mutated

to become malignant. In some, if not all cases, several mutations must

take place for a cell to become malignant.

In recent years, many researchers have described genetic

abnormalities found in malignant cells. These genetic changes consist of

several types of mutation. Some are point mutations that change some

part of the reading frame of a gene. If present in the coding portion of a

peptide, this type of mutation can produce an altered protein. Frequently,

altered proteins fail to perform their customary physiological functions. In some instances this poses a health risk. In some instances, the altered

protein is, itself, a health risk. Alternatively, a point mutation could alter

the behavior of a non-coding region of the genetic material, affecting the

control mechanisms that choreograph the timing of gene expression or the

quantity of material produced. Another type of mutation is a genetic

translocation whereby two strands of DNA become displaced, such that a

portion of part A crosses over to become affixed to part B. In a

translocation event, a whole series of genes can shift to become subject to

control elements that, in normal physiology, control an entirely different set

of genes. This can produce devastating physiological effects. For

example, such effect may arise in genes whose normal function requires

precise control that is lacking in the displaced condition. A third, related,

type of mutation is an alteration in gene amplification, in which the number

of copies of a gene is increased. This, too, can have adverse

consequences.

Appreciation and understanding of the complexity of this process is

limited by an incomplete knowledge of the normal function of these genes

whose function is implicated in the cascade that leads to malignancy. It

does not appear that a single universal genetic alteration is present in all

cancerous cells. Certain genes, such as p53, are implicated in a great

number of cancers, but there are many malignant cells that do not appear

to have any alteration in p53. Multiple possible paths which independently

produce malignant cell. It appears that "oncogenes" (mutated forms of normal gene product) can cause healthy cells to become malignant when

they are inserted into its DNA. Others, called "tumor suppressor genes"

are believed to act by preventing cells from uncontrolled proliferation.

These and other types of alterations may be required to convert a cell to

malignancy.

Tobacco smoke contains numerous compounds that are deposited in

the lungs. As many as 3,000 different chemicals have been identified in

tobacco smoke. It is believed that smoking associated disease are caused

by these chemicals, acting alone or in concert with other chemicals. Some

of these compounds are known carcinogens of the class termed mutagens.

Mutagens are compounds which increase the rate of mutations in exposed

cells. While most mutations are not associated with cancer, some are.

The accumulation of mutations in cells over time increases the chance that

one cell will convert into a state of uncontrolled growth and become an

invasive tumor. A carcinogenic chemical agent may begin a process that

will lead to a cancer in some cell, but there is no disease until the threshold

of cellular conversion to uncontrolled replication is reached.

Some researchers believe the process by which the carcinogens that

are in smoke cause mutations is by generation of free radicals that interfere

with DNA-protein interactions which produce permanent changes in the

DNA. An example of this class of carcinogens is the family of compounds

called nitrosamines. Certain nitrosamines, for example, 4 - (N -

nitrosoamino) - 4- (3 pyridyl) - 1 -butanone, have been reported to be carcinogenic, or procarcinogenic. By procarcinogenic it is meant that

metabolites of the compounds are carcinogenic or of greater carcinogenicity

than the unmetabolized compound. In some instances, free radicals

increase the mutagenic activity of nitrosamines and other substances in

tobacco smoke.

Free radicals are species whose outermost electron is unpaired.

Often free radicals are associated with oxygen, as in an hydroxide or nitric

oxide or superoxide, but they can involve other molecules as well. Usually

free radicals are short lived and highly reactive. When proximate, releasing

the energy of a free radical dislocates interactions of proteins and nucleic

acids in DNA in an active state such as DNA synthesis or repair, and

thereby causes mutation. Superoxide is an O₂ molecule that has an extra

electron: O₂ , hydrogen peroxide is H₂O₂ and the hydroxyl radical is an OH

molecule that dissociated from water and is highly reactive: OH^°.

Oxygen free radicals exert their effect by binding to biological

molecules such as lipids, proteins and DNA. One important reaction caused

by oxygen free radicals is formation of peroxide intermediates in the

unsaturated fatty acid moieties of cell membrane lipids. These oxidized

intermediates can then propagate to interfere with the protein:DNA

interactions that leads to mutation, which can ultimately eventuate in

conversion to malignancy.

Free radicals are ubiquitous and are normally part of many metabolic

processes. Mammals produce natural anti-oxidants to neutralize free radicals. For example, the enzyme catalase converts hydroxide anion and

hydrogen peroxide to water and oxygen. Superoxide dismutase is another

enzyme that converts superoxide to H₂O₂. In addition to enzymes, there

are small molecule anti-oxidants such as glutathione and alpha-tocopherol.

Other potent anti-oxidants commonly used in the food industry include

("BHT") also termed 2,6-bis( 1 , 1 -dimethylethyl)-4-methyl-phenol or

2,6-di-tert-butyl-p-cresol, and butylated hydroxy anisole ("BHA") with the

systematic name ( 1 , 1 -dimethylethyl)-4-methoxy phenol, and propyl gallate

with the systematic name 3,4,5-trihydroxy-benzoic acid, and propyl ester.

Aspects of BHT are discussed in Witschi et al. , "Metabolism and Pulmonary

Toxicity of Butylated Hydroxytoluene (BHT)," Pharmac. Ther., Vol. 42, pp

89-1 1 3 (1 989) the teachings of which are incorporated herein by

reference.

One approach to reducing the long term effect of chronic exposure to

mutagens is regular treatment with anti-oxidants to interfere with free

radicals before they cause damage. Dietary anti-oxidants are available in

many forms, including food supplements and food preservatives.

It is the particular advantage of this present invention to apply an

effective dose of high potency anti-oxidants topically to the lung epithelium

where they directly act on the free radicals present in the lungs.

Another aspect of the present invention concerns retinoids.

Retinoids have also been considered in monodrug antineoplastic therapy

both for the treatment of cancer and for cancer prevention. Retinoids are a family of isoprenoid compounds that include beta-carotene, vitamin A,

retinal, retinoic acid, etc. Dietary beta-carotene is metabolized to produce

two retinol molecules. Retinol is an essential vitamin. Vitamin A circulates

in a protein complex called retinol binding protein. Retinol collects in cells

that have receptors for this complex, most notably epithelial cells. In the

specialized cells of the retina, it is converted to the aldehyde form, retinal,

the visual pigment of the eye. Vitamin A deficiency is associated with a

number of effects, including night blindness, abnormal keratinazation of the

skin, dry skin and conjunctival abnormalities.

Retinoic acid is an intracellular signal molecule that acts directly on a

nuclear receptor to catalyze the expression of a unique sequence of genes.

There are two families of currently identified retinoic acid receptors, RAR

and RXR. Two isomeric forms exist for the double bonds in the retinoid

isoprene chain; cis and trans. Note that cis and trans double bonds can be

at various lications such as 1 3-cis and 9 cis etc. Retinoic acid promotes

cellular differentiation and plays an important role in the timing of

expression in embryonic development. It is known to be teratogenic in

pregnant animals. 1 3-c/s retinoic acid (isotretinoin) is effective topically in

the management of acne through its direct action on keratinocytes. All-

trans retinoic acid (tretinoin), isotretinoin and 9-cis retinoic acid have been

shown to have antiproliferative effects on cultured leukemia cells, as have a

number of synthetic analogs. The antiproliferative effect has been shown to have clinical utility in the management of cancer in certain settings when

used as oral formulations.

Despite successes, other retinoid clinical trials have proven

disappointing. For example, a recently published trial using a retinoic acid

derivative that, in laboratory tests was more potent than the natural

compound, was ineffective in patients who were long term smokers. Kurie

et al., "4-Hydroxyphenylretinamide in Reversal of Bronchial Metaplasia and

Displasia In Smokers," Proceedings of ASCO 1 8:473a (1 999) (American

Society of Clinical Oncology, Atlanta, VA) found no significant effect from

an oral retinoid administration. Parthasarathy et al., "Aerosol delivery of

liposomal all-trans-retinoic acid to the lungs," Cancer Chemotherapy

Pharmacol 43(4):277:277-83 (1 999) notes aerosol delivery of lipsosmal

ATRA as an anti-inflammatory accompanying cancer chemotherapy.

Summary of the Invention

Ingested dietary anti-oxidants distribute throughout the body upon

ingestion and do not reach adequate levels in the lungs. A novel aspect of

the present invention is in the combination of high effect, lipophilic

antioxidants engaged with the lipid medium of the lungs to interdict the

propagation of nuclear mutations. Without being bound by ant particular

theory, hydrophilic anti-oxidants tend to clear more quickly than lipophilic

antioxidants. In addition, hydrophilic antioxidants do not partition in lipids.

This reduces the efficacy of DNA protection afforded by hydrophilic antioxidants by reducing entry into the cell nucleus. In particular

embodiments, high effect, lipophilic antioxidants are delivered in

combination with an antiproliferative agent such as a retinoid.

This invention includes a cigarette filter-type device comprising a

pharmaceutically acceptable high-effect antioxidant, and particularly

antioxidant present in an inhalable therapeutically effective amount. In

particular embodiments antioxidant is present in at least about 0.2 to about

1 .5 milligrams and particularly about 0.5 to 1 milligram. In some

embodiments the device further comprises a pharmaceutically acceptable

volatile compound admixed with the antioxidant and or a therapeutically

effective amount of a pharmaceutically acceptable retinoid such as all-trans

retinoic acid or 1 3-cis retinoic acid.

Noted are high-effect antioxidants selected from the group

comprising BHT, BHA or propyl gallate.

Further included is a particular cigarette filter-type device, which

comprises:

(a) an outer elongate cylinder of tobacco smoke filter material, (b) an inner

elongate cylinder generally coextensive with the outer elongate cylinder of

(a), and the inner elongate cylinder comprising high-effect antioxidant in

powder form in inhalable therapeutically effective amount with the powder

of averaged particle size of less than about 6 u, and in some instances less

than about 2 u. One specific cigarette filter-type device of this invention comprises

an outer elongate cylinder of tobacco smoke filter material said device

further comprising rupturable microcapsules comprising high-effect

antioxidant, optionally in powder form. Noted is rupturable microcapsules

comprising high-effect antioxidant and optionally retinoids such as ATRA.

In some instances the microcapsules further comprise a pharmaceutically

acceptable high-effect antioxidant solvent.

The invention yet further includes an aerosol device comprising a

therapeutically effective amount of a pharmaceutically acceptable high-

effect antioxidant in diluent-propellant. Useful high-effect antioxidant is

selected from the group comprising BHT, BHA or propyl gallate. Some

aerosol devices are metered dose devices or pump type devices. These

devices are usefully configured to deliver aerosol averaged particle sizes of

less than about 6 u, and optionally particles of less than 2 u. A particular

metered dose device uses BHT in an amount of at least about 300 mg in

about 1 5 ml diluent-propellant. Also noted is BHT in an amount of at least

about 30 mg/1 ml diluent-propellant.

This invention also includes a method of treating deep lung surfaces

to provide an lipid peroxidation-mitigating surface by the step of inhaling at

least about .5 mg of high-effect antioxidant, optionally in averaged particle

sizes of about 6 microns ("μ") or less including about 2 μ or less. In one

embodiment the method is accomplished by means of a cigarette filter-type

device. In such method the high-effect antioxidant is selected from the group comprising BHT, BHA or propyl gallate. Alternatively the method is

by means of an aerosol device. In practicing the method in one

embodiment it is contemplated that at least about 1 5% of a subject's

alveolar surface is treated with at least about 1 2 nanograms/m².

This invention further includes a method of creating a lipid

peroxidation-reductive deep lung surface by the process of inhaling at least

about .5 mg of high-effect antioxidant such as BHT, BHA or propyl gallate

and at least about 1 mg retinoid, wherein said antioxidant and retinoid of

averaged particle sizes of about 6μ or less. In one example inhalation is by

means of a cigarette filter-type device, optionally co-timely, coordinated or

coincident with inhalation of tobacco smoke.

Brief Description of the Drawings

Fig 1 . is a diagrammatic representation of a cigarette filter-type

device attached to a cigarette.

Fig 2. is a diagrammatic representation of a cigarette filter-type

device attached to a cigarette.

Fig 3. is a diagrammatic representation of a cigarette filter-type

device with rupturable microcapsules.

Fig. 4. is a diagrammatic representation of a metered dose inhaler.

Fig. 5. is a diagrammatic representation of a pump type inhaler.

Detailed Description of the Invention This invention will be better understood with reference to the

following definitions:

A. "Therapeutically effective amount" as to high-effect antioxidant

administered on a daily basis shall mean at least about 0.5 mg per

5 day. For particular antioxidants, BHT, propyl gallate or BHA about

0.5 to about 1 0 mg and in a particular embodiment about 0.5 to

about 1 .5 mg, and particularly about 1 mg are therapeutically

effective, but further noting about 1 .5 mg to about 3 mg, and further

up to about 5 mg.

lo It is to be understood that therapeutically effective amount doses are

also conveniently administered as divided doses on a "per cigarette" basis

such as by placing antioxidant in a cigarette. As to inhaled smoke or an

air/smoke mixture, about 1 ng of antioxidant /cc of inhaled smoke

(including particulates) is therapeutic, with particular reference to about

is 100 to about 400 ng/cc.

In a multiple use cigarette filter, anticipating at least about 5

cigarettes being smoked daily from about .5 to about 2 ng/cc of inhaled

smoke are therapeutically effective doses.

A cigarette filter type device which contains from about 0.1 to about

20 10 mg of high effect antioxidant is therapeutically effective. This range

has a breadth, in part, due to the variability by which the antioxidant will

mobilize from different filter materials, their compaction and the airflow

during smoking. As to an aerosol or pump type inhaler a delivered dose of about .5 to

about 1 .5 mg, and particularly about 1 mg is therapeutically effective. In a

metered-dose inhaler a therapeutically effective amount is about 1 mg in a

vapor or aerosol volume of from about 0.01 cc to about 0.2 cc. In a

nebulizer providing about 1 to about 5 cc of aerosol, a concentration of

antioxidant of from about 0.2 to about 1 mg/cc is therapeutically effective.

In one embodiment from about 1 to about 5 cc is the volume of liquid put

in the nebulizer. That volume is dispersed in the air by the nebulizer.

Typically the pressure unit produces 3 to 10 liters per minute, and runs for

3 to 1 5 minutes. In this example, this volume of air is about 20 to 80 liters.

B. "Deep lung" shall mean the ends of the bronchial tree, in the region

called the respiratory zone, where the bronchial walls are open to

exchange of gases.

C. High-Effect Antioxidant including BHT, BHA, propyl gallate,

anthocyanins shall mean that the general class of antioxidants

displays at least about twice the anti-oxidant effect measured over

time as alpha-tocopherol. In one embodiment of such a test, a

standard vial of unsaturated phopspholipid and alpha -tocopherol are

maintained at 37^°C. By visual inspection the phospholipid material

will turn brown in about 1 month. An equimolar amount of BHT or

BHA or other high-effect anti-oxidant will keep a standardized

amount of unsaturated phopspholipid from browning for at least

twice as long. Alternatively, free radical scavenging activities can be determined by

a number of methods. Reference is made to Maulik G, et al. , "Evaluation of

antioxidant effectiveness of a few herbal plants," Free Radic Res

Aug;27(2):221 -8 (1 997), the teachings of which are incorporated herein by

reference. One method of determining free radical scavenging is by

chemiluminescence. Peroxyl radical is generated from 2,2'-azobis(2-

amidinopropane) dihydrochloride (AAPH), superoxide radical (O2-) from

xanthine/xanthine oxidase (XO) and hydroxyl radical (OH) from

Xanthine/XO/FeCI3/ EDTA. In addition, 02- and OH. scavenging activities

are determined by cytochrome C reduction and deoxyribose oxidation

methods, respectively.

There are also many pharmaceutically acceptable high-effect

anti-oxidants including plant extracts, water soluble anti-oxidants include

vitamin C, sorbates, citric acid, thioesters and other sulfur reducing agents,

such as methionine, taurine, cysteine and glutathione. Lipophilic

anti-oxidants include both natural and synthetic compositions. The

carotenoids and tocopherols are of lesser effect. Another category of high-

effect antioxidant is the isoflavone derivatives, such as Bachanin A. There

are also phenols/O-methyl phenols including BHA, BHT, propyl gallate,

tertiary butyl hydroquinone, curcumin, eugenol, and ferulic acid. Also

noted are salen-metal complexes such as those described in US 5,696, 1 09

to Malfroy et al., the teachings of which are incorporated herein by

reference. D. "Aerosol device" shall be broadly understood to encompass

pressurized and non-pressurized (spray) devices either of which may

include spacers, and electrohydrodynamic devices. These devices

include metered-dose inhalers, atomizers, piezo electric aerosol

generators, powder blowers or powder insufflators, vaporizers and

nebulizers. A number of aerosol devices are described in

Pharmaceutical Dosage Forms and Drug Delivery Systems. Ansel et

al., Sixth Ed. (Williams & Wilkins, Media Penn., 1 995) as well as

Ranucci et al, "Effect of Actuator Design on Metered-Dose Inhaler

Plume Particle Size" Pharmacology Technology 1 6(3):84-92, ( 1 992),

the teachings of which are incorporated herein by reference.

In practice pressurized aerosol devices are pressurized from about 25

to about 55 pounds per square inch gauge (psig). This invention is not to

be limited by the device used to provide inhalable high effect antioxidants.

Of the many suitable devices, mention is made of pump inhalers, metered-

dose inhalers, and electrohydrodynamic aerosol generators and other

devices are also conveniently employed with enhancers such as spacers

and the like. Note is made of the metered dose inhalers marked under the

following names: Tidal Inhaler™ from Fisons, Azmacort™ from Rhδne-

Poulenc Rorer, Beclovent™ and Flovent™ from Glaxo Wellcome Airomir™

from 3M Healthcare, and dry powder inhalers such as Pulmicort

Turbohaler™ from Astra and Severent Diskkhaler™ from Glaxo Wellcome.

Also noted are electrohydrodynamic aerosol devices such as are described in US 5,91 1 5,377, US 5,81 3,61 4 and US 5,655,51 7 the teachings of

which are incorporated herein by reference. Aerosol devices are also

employed with diluent-propellants which encompass respiratory aerosol

delivery adjuncts including surface-active agents, solvents, propellants such

as liquified gas or a mixture of liquified gases {e.g., chloro-fluorocarbons,

dichlorodifluromethane, dichlorotetrafluroethane, trichloromonofluroethane,

octafluorocyclobutane), non liquified compressed gases (e.g., carbon

dioxide, nitrogen, nitrous oxide) and diluents and solvent such as water and

ethyl alcohol. Two- and three-phase systems are noted as well as

compressed gas.

Aerosols produced by pressurized metered dose inhalers generally

conform to "log-normal" functions. Aerosols are produced by many

devices including a piezo-electric device that provides an ultrasonic

vibration and devices that deliver dry powder aerosols. Drug delivery

properties of log-normal functions are frequently characterized by mass

mean aerodynamic diameter (MMAD) and geometric standard deviation

(GSD) values. Note is made of Raabe, "Particle Size Analysis Utilizing

Grouped Data and Log-Normal Distribution," J. Aerosol. Sci, 2:289-303

( 1 971 ), Martonen et al., "Deposition Patterns of Aerosolized Drugs Within

Human Lungs: Effects of Ventilatory Parameters," Pharmaceutical

Research, 1 0(6):871 -878 (1 993), and Martonen et al., "Use of Analytically

Defined Estimates of Aerosol Respirable Fraction to Predict Lung Deposition Patterns," Pharmaceutical Research. 9(1 2): 1 634-1 639 (1 992), the

teachings of which are incorporated herein by reference.

E. "Co-timely" shall mean administration of a high-effect antioxidant

while tobacco smoke is being inspired.

F. "Coordinated" refers to mean pre- and post- dosing via inhaler or the

like. "Coordinated" in the practice of the present invention refers to

administration of a high-effect antioxidant in a therapeutically

effective amount within at least about 1 to 30, or about 60 to 1 20

minutes before/after smoking a tobacco cigarette, and particularly

about 5 minutes or less prior to cigarette smoking

G. "Coincident" refers to a combination drug therapy with a retinoid and

a high-effect antioxidant. Coincident means dosing comprising

administration of a retinoid while a high-effect antioxidant is present

in at least about a therapeutic amount on the lung surface (or "in

delivery" to the lung surface), and potentially in higher

concentrations. In a preferred embodiment of coincident

administration, both drugs are administered in a single aerosol.

H. "Cigarette filter-type device" shall mean a device affixed or affixable

to the downstream end of a cigarette. In the usual case the

downstream end of a cigarette is the end nearer the mouth of a

smoker, with the cigarette filter-type device being inserted directly

into the lips or mouth of the smoker or via a cigarette holder. The

term is broadly construed to include filter-type devices which include filamentous materials typically employed to remove tars and other

substances from smoke, but also to include the addition of drugs,

flavorant and the like to smoke even when no substance is removed

from the smoke. Further the term contemplates such devices as

affixed to cigarettes when manufactured forming a single unit, such

devices as provided after cigarette manufacture and affixed later

such as when smoking the cigarette begins, and the device as

inserted or contained within a cigarette holder, in which the cigarette

is inserted prior to smoking the cigarette. Cigarette holders are

generally characterized as having mouthpieces, often similar to those

found on pipes.

I. "Cigarette" is a term to be broadly understood to include smoked

tobacco type devices such as typical paper wrapped cigarettes, as

well as plant leaf or otherwise wrapped cigarettes, and further

including cigars, pipes, hookahs and the like. It is also understood to

mean, generally, a reference to smoked tobacco containing items,

but other smoke producing products for purposeful smoke inhalation

such as marijuana are also contemplated within the term.

J. "Inhalable" is a term referencing the amount of drug within a filter

that will be inhaled in at least about 500cc of inhaled air and or

smoke in the process of smoking a cigarette.

K. "Retinoid" shall mean natural and synthetic derivatives of vitamin A

(retinol). Isotretinoin and tretinoin represent two naturally occurring isomers of retinoic acid. Many other synthetic analogs have also

been studied, including etretinate. Tong, et al., WO 97/3974 "Use

of Inhaled Retinoids in the Prevention of Cancer," the teachings of

which are incorporated herein by reference. Without being bound

by any particular theory it is believed that retinoic acid activates

receptors in cell nuclei, notably RAR and RXR receptors which can

result in significant physiological changes. Such changes include

anti-proliferative effects and/or differentiation. Other retinoids may

require processing by elements in the cytosol before they become

active in changing the pattern of transcription.

Without being bound by any particular theory, it is believed that

retinoids act by inducing differentiation. A variety of in vitro studies have

demonstrated the specialized ability of retinoids to induce terminal

differentiation and inhibit growth of several human leukemia cell lines and

bone marrow cultures obtained from patients with acute myelogenous

leukemia. These studies and many others done in murine cell lines provide

a scientific foundation for the in vivo use of retinoids to treat certain types

of leukemia. Data suggest that retinoids induce growing cells to mature to

become end-stage cells that are no longer capable of dividing.

This property has been tested in clinical trials. Clinical activity of

retinoids has been demonstrated in many diverse premalignant conditions

including squamous cell carcinomas of the skin, basal cell carcinomas,

cutaneous T-cell lymphomas, and leukoplakia. Patients who have demonstrable premalignant lesions and are treated with retinoids are less

likely to develop malignancies, compared to patients who were not treated

with retinoids.

L. "Therapeutically effective" as to retinoid doses will vary with the

presenting condition and subject.

In practice, dosing followed by lung biopsy offers an empirical end

point for retinoid dosing and is within the skill of the medical practitioner.

As a general guide, attention is drawn to doses of about .1 to about 20 mg

total delivered retinoid dose and in a particular embodiment about 0.25 to

about 10 mg, and more particularly about .5 to about 5 mg, as well as

from about 1 0 to about 1 5 mg.

Daily total retinoid dosages such ad ATRA of from about 0.1 to

about 1 5 mg/day are contemplated. Particular reference is made to daily

doses of up to about 6 mg.

It is to be understood that total daily doses are variable. Many

factors including smoking or inhalation "style," amounts of therapeutic

agent per cigarette, and the number of cigarettes smoked effect total daily

dosage. It is contemplated that a health professional will monitor a subject

for symptoms or indicia of hypervitaminosis or build up of therapeutic agent

and reduce (or increase) doses accordingly.

Representative Devices

Fig 1 . is a diagrammatic representation of a cigarette filter-type

device (4) attached to a tobacco containing segment (2) of a cigarette (1 0). Tobacco is noted as (8) and a filamentous filter material is noted as (6). In

some embodiments the high-effect antioxidant or anti-mutagen comprises a

coating on the filter material or a powder intermingled with the filter

material.

Fig 2. is a diagrammatic representation of a cigarette filter-type

device (24) attached to a tobacco containing segment (22) of a cigarette

(20) . Tobacco is noted as (28) and a filamentous filter material is noted as

(26). An additional element of the cigarette filter-type device (24) is high-

effect antioxidant in powder form (27) in carrying tube (23) in air or smoke

flow connection with abutting tobacco containing segment (22). Porous

filter material plugs (21 ) contain the powder (27) in tube (23).

Fig 3. is a diagrammatic representation of a cigarette filter-type

device (34) with rupturable microcapsules (38) containing high effect

antioxidant and or anti-mutagens dispersed amidst filamentous filter (36).

A tobacco containing element is shown in phantom (32).

Fig. 4. is a diagrammatic representation of a metered dose inhaler

(40) . Holder (42) supports a pressurized container (46) comprising

antioxidant and propellant in connection with metered delivery element

(44) . Upon downward pressure on the pressurized container (46) metered

delivery element (44) delivers an aerosol mist directed through opening

(48).

Fig. 5. is a diagrammatic representation of a pump type inhaler (50) .

Container (52) holds a solution or suspension (54) comprising high effect antioxidant and solvent/carrier liquid propellant in connection with pump

means (58) in connection with atomizer head (56).

Methodological Aspects

Dosing smokers with appropriate amounts of high-effect antioxidant

requires particular attention. It is appreciated that in specific instances,

dosing subjects with compromised respiration or lung elasticity or function

will necessarily deviate from preferred ranges. In some instances, modified

inspiratory regimens are required. With aerosolizing devices, multiple

inhalations are required by certain individuals to dose bronchiole or alveolar

surfaces to a specific level. In some instances, positive pressure ventilation

is required to introduce high-effect anti-oxidant into the lungs of

compromised subjects or to introduce dosages in controlled amounts or to

specific depths.

Smaller alveoli produce lung surfactant which coats the inside of the

sac, reducing surface tension and stabilizing them. Lung surfactant is a

mixture of several apoproteins and phospholipids. It is produced by type 2

cells of the alveoli, and is secreted into the lumen. There is an active

recycling process whereby surfactant is absorbed and its components

reassembled into surfactant for secretion. This lining of surfactant is a

target for oxidative damage by inhaled free radicals. Lipid peroxides

produced by oxidation can propagate the damage inside the cells lining the

airways, sometimes causing mutations. The lining surfactant is also the

target for lipophilic anti-oxidants. The surfactant provides a bed to sustain levels of high effect anti-oxidant in the milieu where they can maximize

their benefit.

It is to be understood that in some subjects, often associated with

respiratory pathologies, a physiological dead space is present distinct from

the anatomic dead space. Certain alveoli will be either over or under

ventilated under "at rest" conditions. The volume of gas in non-perfused

alveoli and any volume of air in the alveoli in excess of that necessary to

arteriolize the blood in the alveolar capillaries is part of the dead space (non-

equilibrating) gas volume. Without being bound by any particular theory, in

the instant invention it is thought that an anti-oxidant such as BHA or a

combination of BHT and BHA will eliminate or neutralize some free radicals

and reduce the mutation rate in the lungs. By reducing the overall mutation

rate the user has a reduced incidence of or delayed development of lung

cancer.

Delivery of the anti-mutagenic therapy of this invention is usefully in

aerosol form. Aerosol form refers to the condition of the inspired drug

upon exiting the aerosol device. It is contemplated that a spray in aerosol

form encompasses aqueous and oleaginous droplets. In some

embodiments aerosol form is a dry powder. In some embodiments the

aerosol form begins as a dry powder and in some the aerosol begins as a

droplet of drug and volatile carrier, wherein the carrier evaporates leaving a

dry powder. In yet other forms, the spray is droplets containing drug

particle or liposomes containing drug. Noted volatile carriers are lower alkyl alcohols such as ethanol and isobutanol. When used withing filter type

devices volatile carriers are usefully contained in rupturable microcapsules

Powders for inhaled administration are also contemplated. In some

embodiments powders are dusted directly onto filter material. These may

be prepared by any of a number of methods well known in the art. By way

of example, a useful preparation is similar to that for cromolyn sodium.

The therapeutic is compacted with lactose in particle size of about 30-60 μ,

while the cromolyn is from about 1 to aboutl O μ. The capsule of lactose

and drug is broken capsule and placed in inhaler a powder, e.g. Spinhaler™

from Fisons. Upon inhalation, the larger lactose particles do not proceed

beyond the upper airways while the smaller drug particles are inspired into

the deeper lung. Reference is made to Ranucci et al., "Phase Doppler

Anemometry: A Technique for Determining Aerosol Plume-Particle Size and

Velocity, " Pharmaceutical Technology, 1 7:62-73 ( 1 993) the teachings of

which are incorporated herein by reference.

In one embodiment, this invention comprises is a product that is

inhaled to cover the surface of the lungs, and optionally accompanied by

printed and pictorial instructions for use. Inhalation raises the levels of

these nutritional and/or anti-oxidant supplements to an effective level in and

on the target tissue. In a specific embodiment the drug is administered

once a day from a device that is inexpensive to manufacture, and is

discarded after a few uses. A related aspect of the present invention is to employ the therapeutic

agents of the persent invention in "smokeless" tobacco products such as

snuff and chewing tobacco. In such instances the therapeutic effect as to

the surface of the oral cavity is noted.

One particular embodiment of the therapeutic device of the present

invention comprises a means for delivery of a combination of an anti¬

oxidant and retinoid (e.g., retinoic) to the lungs of a subject. Attention is

drawn to "at risk" subjects. Particularly "at risk" subjects are tobacco

smokers, ex-smokers, subjects with long term exposure to such hazards as

asbestos and hay, as well as miners of coal, uranium, beryllium and the

like.

Cigarette Filter-Type Devices

Fibre- or filament-containing filters for cigarettes are well known. In

one form of construction, the filter body consists of a tow of continuous

filaments, commonly cellulose acetate (acetate) filaments, arranged parallel

to the long axis of the cigarette. In another form of construction, the filter

body consists of pleated or fluted paper compressed into a cylinder. Such

forms of construction contain a single filter element and may be called

"mono" filters. Another form of construction is the so-called "dual" filter

which contains two filter elements, for example a paper filter towards the

interior and a tow filter towards the exterior of the cigarette. A further

form of construction is the so-called "triple" filter, which resembles a dual

filter except that a quantity of a third substance such as activated carbon is interposed between the two filter elements hereinbefore mentioned.

Attention is drawn to US 5,839,448 to Woodings, the teachings of which

are incorporate herein by reference.

In some cigarette filter-type devices active or aromatic substances

are introduced from the device into the smoke inhaled from a cigarette in

order to modify its taste or to confer additional properties. For example,

U.S. Pat. No. 3,991 ,773 and U.S. Pat. No. 3,635,226 describe

mechanisms for humidifying cigarette smoke by including rupturable

capsules or micro-capsules filled with a liquid, within the filter of the

cigarette. The smoker obtains the humidification effect by squeezing the

filter to rupture the capsules prior to smoking. In some instances, liquid

incorporated in the capsules contains flavorings or a synthetic saliva

solution, for example. U.S. Pat. No. 4,498,485 discloses the inclusion of

interferon into the tobacco of a cigarette for administering it directly to the

lungs of the smoker. Note is made of US 5,472,002 to Covarrubias the

teachings of which are incorporate herein by reference.

Example 1

A BHA mixture for application to the filter material is compounded as

follows:

BHA 1 mg is dissolved in 1 cc of ethanol. The solution is applied to a

conventional cellulose acetate cigarette filter and air dried. The filter is

then affixed to the end of a tobacco cigarette. Example 2

A 24 year old female smoker is provided with a filter cigarette of

Example 1 . She smokes the cigarette in her normal manner inhaling the

tobacco smoke. In the course of smoking the cigarette she inhales at least

about .5 mg of BHA.

Example 3

A BHT mixture for application to the filter material is compounded as

follows:

BHT, 0.8mg is dissolved in .5 cc of ethanol. The solution is applied

to a conventional cellulose acetate cigarette filter and air dried. The filter is

then affixed to the end of a tobacco cigarette.

Example 4

A 67 year old male smoker is provided with a filter cigarette of

Example 3. He smokes the cigarette in his normal manner inhaling the

tobacco smoke. In the course of smoking the cigarette he inhales at least

about .4 mg of BHT.

Example 5

A BHT-ATRA mixture for application to the filter material is

compounded as follows: BHT 1 mg and .5 mg ATRA are dissolved in 1 cc of ethanol. The

solution is applied to a conventional cellulose acetate cigarette filter and air

dried. The filter is then affixed to the end of a tobacco cigarette.

Example 6

A 35 year old male smoker is provided with the cigarette of Example

5. He smokes the cigarette in his normal manner inhaling the tobacco

smoke. In the course of smoking the cigarette he inhales at least about .4

mg of BHT and at least about .2 mg ATRA.

Example 7

A BHT-ATRA mixture for application to the filter material is

compounded as follows:

BHT 1 mg and .5 mg ATRA are encapsulated in rupturable

microcapsules each containing about 0.1 mg of the mixture. The

rupturable microcapsules are located in cigarette filter-type material and the

filter affixed to a cigarette.

Example 8

An 1 8 year old male smoker is provided with the cigarette of

Example 7. With thumb and forefinger pressure on the cigarette filter he

ruptures the microcapsules within the filter and their contents is distributed

within the filter material. Promptly thereafter he smokes the cigarette in his normal manner inhaling the tobacco smoke. In the course of smoking the

cigarette he inhales at least about .5 mg of BHT and 0.4 mg of ATRA

creating an lipid peroxidation-reductive deep lung surface.

Example 9

A BHT mixture for application to the filter material is compounded as

follows:

BHT 1 mg is encapsulated in rupturable microcapsules each

containing about 0.1 mg BHT. The rupturable microcapsules are located in

cigarette filter-type material and the filter affixed to a cigarette.

Example 1 0

A 35 year old male smoker is provided with the cigarette of Example

9. With thumb and forefinger pressure on the cigarette filter he ruptures

the microcapsules within the filter and their contents is distributed within

the filter material. Promptly thereafter he smokes the cigarette in his

normal manner inhaling the tobacco smoke. In the course of smoking the

cigarette he inhales at least about .5 mg of BHT.

All publications mentioned herein above are hereby incorporated in

their entirety by reference

Claims

1 . A cigarette filter-type device comprising a pharmaceutically acceptable

high-effect antioxidant.

2. The device of Claim 1 wherein said antioxidant is present in an inhalable

therapeutically effective amount.

3. The device of Claim 1 wherein said antioxidant is present in at least

about 0.2 to about 1 gram.

4. The device of Claim 1 wherein further comprising a pharmaceutically

acceptable volatile compound admixed with said antioxidant.

5. The device of Claim 1 wherein said high-effect antioxidant is selected

from the group comprising BHT, BHA or propyl gallate.

6. A cigarette filter-type device, which comprises:

(a) an outer elongate cylinder of tobacco smoke filter material,

(b) an inner elongate cylinder

generally coextensive with the outer elongate cylinder of (a),

said inner elongate cylinder comprising high-effect antioxidant in

powder form in inhalable therapeutically effective amount, and

(c) said powder of averaged particle size of less than about 6 u.

7. The device of Claim 6 wherein said high-effect antioxidant is selected

from the group comprising BHT, BHA or propyl gallate.

8. A cigarette filter-type device, which comprises an outer elongate

cylinder of tobacco smoke filter material said device further comprising

rupturable microcapsules comprising high-effect antioxidant.

9. The high-effect antioxidant of Claim 8 in powder form.

10. Rupturable microcapsules comprising high-effect antioxidant

1 1 . The microcapsules of Claim 1 1 further comprising a pharmaceutically

acceptable high-effect antioxidant solvent.

1 2. An aerosol device comprising a therapeutically effective amount of a

pharmaceutically acceptable high-effect antioxidant in diluent-propellant.

1 3. The device of Claim 1 2 wherein said high-effect antioxidant is selected

from the group comprising BHT, BHA or propyl gallate.

14. The device of Claim 1 2 wherein said device is a metered dose device.

1 5. The device of Claim 1 2 wherein said device is a pump-type device

1 6. The device of Claim 1 further comprising a therapeutically effective

amount of a pharmaceutically acceptable retinoid.

1 7. The device of Claim 1 6 wherein the retinoid is all-transretinoic acid or

1 3-cis retinoic acid.

1 8. A method of treating deep lung surfaces to provide an lipid

peroxidation-mitigating surface by the step of inhaling at least about .5 mg

of high-effect antioxidant.

1 9. The method of Claim 1 8 wherein said antioxidant is in averaged

particle sizes of about 6 μ or less.

20. The method of Claim 1 8 wherein inhalation is by means of a cigarette

filter-type device.

21 . The method of Claim 1 8 wherein said high-effect antioxidant is

selected from the group comprising BHT, BHA or propyl gallate.

23. The method of Claim 1 8 wherein inhalation is by means of an aerosol

device.

24. The method of Claim 18 wherein at least about 1 5% of said alveolar

surface is treated with at least about 1 2 nanograms/m².

25. A method of creating an lipid peroxidation-reductive deep lung surface

by the process of

inhaling at least about .5 mg of high-effect antioxidant and at least

about .1 mg retinoid, wherein said antioxidant and retinoid of

averaged particle sizes of about 6 μ or less.

26. The method of Claim 25 wherein said high-effect antioxidant is

selected from the group comprising BHT, BHA or propyl gallate.

27. The method of Claim 1 8 wherein inhalation is by means of a cigarette

filter-type device.

28. The method of Claim 1 8 wherein said high-effect antioxidant is

selected from the group comprising BHT, BHA or propyl gallate.

29. The method of Claim 27 wherein said inhalation is co-timely with

inhalation of tobacco smoke.

30. The method of Claim 27 wherein said inhalation is coordinated with

inhalation of tobacco smoke.

31 . The method of Claim 27 wherein said inhalation is coincident with

inhalation of tobacco smoke.