Methods for prolongation of climax time in a patient in need thereof are presented, as are methods for treating premature ejaculation by local administration of a Clostridial neurotoxin, such as botulinum neurotoxin, to the patient, are provided.

Methods for treating premature ejaculation are presented. More particularly and in one aspect, methods for treating premature ejaculation by administration of a neurotoxin, such a botulinum neurotoxin, to a patient are provided.
Premature ejaculation (PE) is a common sexual dysfunction in men, particularly those in the age range of about 18 to about 40 years old. Premature ejaculation can be generally defined as the occurrence of ejaculation prior to or sooner than hoped for by one or both sexual partners [e.g. see 'The Merck Manual', 16th Edition, p 1576, published by Merck Research Laboratories, 1992]. As one example, premature ejaculation can be experienced as ejaculation before, upon or shortly after penile penetration of a sexual partner. If the instances of premature ejaculation are few and far between, then such occurrences may not be a cause for concern. However, if instances of premature ejaculation occur practically every time intercourse is attempted, or even if premature ejaculation occurs even greater than about 10% or about 20% of the time intercourse is attempted, then treatment of the condition is likely to be warranted.
Premature ejaculation may be classified as primary or secondary, in accordance with the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV), which classifies sexual disorders into 4 particular categories: (1) primary, (2) general medical condition-related, (3) substance-induced, and (4) not otherwise specified. Primary applies to individuals who have had the condition since they became capable of functioning sexually (i.e., postpuberty). Secondary indicates that the condition manifests itself in an individual where an acceptable level of ejaculatory control was previously had, and then began to experiencing premature ejaculation thereafter. With secondary premature ejaculation, the problem does not relate to a general medical disorder, and it is usually not related to substance inducement, however, hyperexcitability might in particular instances relate to psychotropic drug use and resolves when the drug is withdrawn.
The prevalence rate of premature ejaculation in American males is estimated to range from 30-70%. The National Health and Social Life Survey (NHSLS) indicates a prevalence of 30%, which is fairly steady through all adult age categories. Premature ejaculation can occur at virtually any age in an adult man's life. As a reported condition, it is most common in younger men (aged 18-30 years old) but may also occur in conjunction with secondary impotence in men aged 45-65 years.
Compositions that can be utilized for treating premature ejaculation are known. For example, use of selective serotonin reuptake inhibitors (SSRI's) for treating premature ejaculation are known and claimed, for example, in U.S. Pat. No. 7,105,516 which is directed to novel SSRI's effective for the treatment of premature ejaculation, as well as U.S. Pat. No. 6,777,437. Additional methods for treating premature ejaculation can be found in U.S. Pat. No. 6,974,839, which teaches administration of an effective amount of a tramadol (a monoamine uptake inhibitor) material to a male prior to sexual intercourse, and in U.S. Pat. No. 6,495,154, which claims delaying the onset of ejaculation in a male by systemically administering to the individual a rapid-release pharmaceutical formulation containing clomipramine and pharmacologically acceptable acid addition salts thereof. U.S. Pat. No. 7,018,648 is directed to a transdermal device for administering testosterone and/or at least one derivative thereof to treat premature ejaculation; U.S. Pat. No. 6,593,335 is directed to method of treating premature ejaculation by administration of a potassium channel opener; U.S. Pat. No. 6,727,283 discloses oral ingestion of an essentially nonaqueous, liquid concentrate of sertraline hydrochloride. In some examples, fluoxetine or paroxetine, (20 mg and 40 mg, respectively, and taken daily) are also prescribed in order to treat premature ejaculation. Other approaches that are known include application of topical anesthetics, such as lidocaine 5% cream, for example, to the penis before intercourse. Drawbacks associated with the use such anesthetics include undesired reduction in sensitivity and/or short term inability of the patient to achieve an erection.
The anaerobic, gram positive bacterium Clostridium botulinum produces a potent polypeptide neurotoxin, botulinum neurotoxin, which causes a neuroparalytic illness in humans and animals referred to as botulism, however this neurotoxin has now been utilized for decades for treating various conditions in human beings. One unit (U) of botulinum toxin is defined as the LD50 upon intraperitoneal injection into female Swiss Webster mice weighing 18-20 grams each. In other words, one unit of botulinum toxin is the amount of botulinum toxin that kills 50% of a group of female Swiss Webster mice. Seven generally immunologically distinct botulinum neurotoxins have been characterized, these being respectively botulinum neurotoxin serotypes A, B, C1, D, E, F, and G, each of which is distinguished by neutralization with type-specific antibodies. The different serotypes of botulinum toxin vary in the animal species that they affect and in the severity and duration of the paralysis they evoke. For example, it has been determined that botulinum toxin type A is 500 times more potent, as measured by the rate of paralysis produced in the rat, than is botulinum toxin type B. Additionally, botulinum toxin type B has been determined to be non-toxic in primates at a dose of 480 U/kg which is about 12 times the primate LD50 for botulinum toxin type A. The botulinum toxins apparently bind with high affinity to cholinergic motor neurons, are translocated into the neuron and block the presynaptic release of acetylcholine.
Botulinum toxins have been used in clinical settings for the treatment of neuromuscular disorders characterized by hyperactive skeletal muscles. Botulinum toxin type A was approved by the U.S. Food and Drug Administration in 1989 for the treatment of essential blepharospasm, strabismus and hemifacial spasm in patients over the age of twelve. In 2000 the FDA approved commercial preparations of type A and type B botulinum toxin serotypes for the treatment of cervical dystonia, and in 2002 the FDA approved a type A botulinum toxin for the cosmetic treatment of certain hyperkinetic (glabellar) facial wrinkles. Clinical effects of peripheral intramuscular botulinum toxin type A are usually seen within one week of injection and sometimes within a few hours. The typical duration of flaccid muscle paralysis from a single intramuscular injection of botulinum toxin type A can be about three months, although in some cases the effects of a botulinum toxin induced denervation of a gland, such as a salivary gland, have been reported to last for several years.
Although all the botulinum toxins serotypes apparently inhibit release of the neurotransmitter acetylcholine at the neuromuscular junction, they do so by affecting different neurosecretory proteins and/or cleaving these proteins at different sites. Botulinum toxin A is a zinc endopeptidase which can specifically hydrolyze a peptide linkage of the intracellular, vesicle associated protein SNAP-25. Botulinum type E also cleaves the 25 kiloDalton (kD) synaptosomal associated protein (SNAP-25), but targets different amino acid sequences within this protein, as compared to botulinum toxin type A. Botulinum toxin types B, D, F and G act on vesicle-associated protein (VAMP, also called synaptobrevin), with each serotype cleaving the protein at a different site. Finally, botulinum toxin type C1 has been shown to cleave both syntaxin and SNAP-25.
Regardless of serotype, the molecular mechanism of toxin intoxication appears to be similar and to involve at least three steps or stages. In the first step of the process, the toxin binds to the presynaptic membrane of the target neuron through a specific interaction between the heavy chain (H chain) and a cell surface receptor; the receptor is thought to be different for each serotype of botulinum toxin and for tetanus toxin. The carboxyl end segment of the H chain, Hc, appears to be important for targeting of the toxin to the cell surface. In the second step, the toxin crosses the plasma membrane of the poisoned cell. The toxin is first engulfed by the cell through receptor-mediated endocytosis, and an endosome containing the toxin is formed. The toxin then escapes the endosome into the cytoplasm of the cell. This last step is thought to be mediated by the amino end segment of the H chain, HN, which triggers a conformational change of the toxin in response to a pH of about 5.5 or lower. Endosomes are known to possess a proton pump which decreases intra endosomal pH. The conformational shift exposes hydrophobic residues in the toxin, which permits the toxin to embed itself in the endosomal membrane. The toxin then translocates through the endosomal membrane into the cytosol. The last step of the mechanism of botulinum toxin activity appears to involve reduction of the disulfide bond joining the H and L chain. The entire toxic activity of botulinum and tetanus toxins is contained in the L chain of the holotoxin; the L chain is a zinc (Zn++) endopeptidase which selectively cleaves proteins essential for recognition and docking of neurotransmitter-containing vesicles with the cytoplasmic surface of the plasma membrane, and fusion of the vesicles with the plasma membrane. Tetanus neurotoxin, botulinum toxin B, D, F, and G cause degradation of synaptobrevin (also called vesicle-associated membrane protein (VAMP)), a synaptosomal membrane protein. Most of the VAMP present at the cytosolic surface of the synaptic vesicle is removed as a result of any one of these cleavage events. Each toxin specifically cleaves a different bond.
The molecular weight of the botulinum toxin protein molecule, for all seven of the known botulinum toxin serotypes, is about 150 kDa. Interestingly, the botulinum toxins are released by Clostridial bacterium as complexes comprising the 150 kD botulinum toxin protein molecule along with associated non-toxin proteins. Thus, the botulinum toxin type A complex can be produced by Clostridial bacterium as 900 kDa, 500 kDa and 300 kDa forms. Botulinum toxin types B and C1 are apparently produced as only a 500 kDa complex. Botulinum toxin type D is produced as both 300 kDa and 500 kDa complexes. Finally, botulinum toxin types E and F are produced as only approximately 300 kDa complexes. The complexes (i.e. molecular weight greater than about 150 kDa) are believed to contain a non-toxin hemagglutinin protein and a non-toxin and non-toxic nonhemagglutinin protein. These two non-toxin proteins (which along with the botulinum toxin molecule can comprise the relevant neurotoxin complex) may act to provide stability against denaturation to the botulinum toxin molecule and protection against digestive acids when toxin is ingested. Additionally, it is possible that the larger (greater than about 150 kDa molecular weight) botulinum toxin complexes may result in a slower rate of diffusion of the botulinum toxin away from a site of intramuscular injection of a botulinum toxin complex. The toxin complexes can be dissociated into toxin protein and hemagglutinin proteins by treating the complex with red blood cells at pH 7.3. The toxin protein has a marked instability upon removal of the hemagglutinin protein.
All the botulinum toxin serotypes are made by Clostridium botulinum bacteria as inactive single chain proteins which must be cleaved or nicked by proteases to become neuroactive. The bacterial strains that make botulinum toxin serotypes A and G possess endogenous proteases and serotypes A and G can therefore be recovered from bacterial cultures in predominantly their active form. In contrast, botulinum toxin serotypes C1, D, and E are synthesized by nonproteolytic strains and are therefore typically unactivated when recovered from culture. Serotypes B and F are produced by both proteolytic and nonproteolytic strains and therefore can be recovered in either the active or inactive form. However, even the proteolytic strains that produce, for example, the botulinum toxin type B serotype only cleave a portion of the toxin produced. The exact proportion of nicked to unnicked molecules depends on the length of incubation and the temperature of the culture. Therefore, a certain percentage of any preparation of, for example, the botulinum toxin type B toxin is likely to be inactive, possibly accounting for a lower potency of botulinum toxin type B as compared to botulinum toxin type A. The presence of inactive botulinum toxin molecules in a clinical preparation will contribute to the overall protein load of the preparation, which has been linked to increased antigenicity, without contributing to its clinical efficacy.
In vitro studies have indicated that botulinum toxin inhibits potassium cation induced release of both acetylcholine and norepinephrine from primary cell cultures of brainstem tissue. Additionally, it has been reported that botulinum toxin inhibits the evoked release of both glycine and glutamate in primary cultures of spinal cord neurons and that in brain synaptosome preparations botulinum toxin inhibits the release of each of the neurotransmitters acetylcholine, dopamine, norepinephrine, CGRP and glutamate.
High quality crystalline botulinum toxin type A can be produced from the Hall A strain of Clostridium botulinum with characteristics of ≧3×107 U/mg, an A260/A278 of less than 0.60 and a distinct pattern of banding on gel electrophoresis. The known Shantz process can be used to obtain crystalline botulinum toxin type A, as set forth in Shantz, E. J., et al, Properties and use of Botulinum toxin and Other Microbial Neurotoxins in Medicine, Microbiol Rev. 56: 80-99 (1992). Generally, the botulinum toxin type A complex can be isolated and purified from an anaerobic fermentation by cultivating Clostridium botulinum type A in a suitable medium. Raw toxin can be harvested by precipitation with sulfuric acid and concentrated by ultramicrofiltration. Purification can be carried out by dissolving the acid precipitate in calcium chloride. The toxin can then be precipitated with cold ethanol. The precipitate can be dissolved in sodium phosphate buffer and centrifuged. Upon drying there can then be obtained approximately 900 kDa crystalline botulinum toxin type A complex with a specific potency of 3×107 LD50 U/mg or greater. This known process can also be used, upon separation out of the non-toxin proteins, to obtain pure botulinum toxins, such as for example: purified botulinum toxin type A with an approximately 150 kDa molecular weight with a specific potency of 1-2×108 LD50 U/mg or greater; purified botulinum toxin type B with an approximately 156 kDa molecular weight with a specific potency of 1-2×108 LD50 U/mg or greater, and; purified botulinum toxin type F with an approximately 155 kDa molecular weight with a specific potency of 1-2×107 LD50 U/mg or greater.
Already prepared and purified botulinum toxins and toxin complexes suitable for preparing pharmaceutical formulations can be obtained from List Biological Laboratories, Inc., Campbell, Calif.; the Centre for Applied Microbiology and Research, Porton Down, U.K.; Wako (Osaka, Japan), as well as from Sigma Chemicals of St Louis, Mo.
It has been reported that a botulinum toxin has been used in clinical settings as follows:
(1) about 75-125 units of BOTOX® per intramuscular injection (multiple muscles) to treat cervical dystonia;
(2) 5-10 units of BOTOX® per intramuscular injection to treat glabellar lines (brow furrows) (5 units injected intramuscularly into the procerus muscle and 10 units injected intramuscularly into each corrugator supercilii muscle);
(3) about 30-80 units of BOTOX® to treat constipation by intrasphincter injection of the puborectalis muscle;
(4) about 1-5 units per muscle of intramuscularly injected BOTOX® to treat blepharospasm by injecting the lateral pre-tarsal orbicularis oculi muscle of the upper lid and the lateral pre-tarsal orbicularis oculi of the lower lid;
(5) to treat strabismus, extraocular muscles have been injected intramuscularly with between about 1-5 units of BOTOX®, the amount injected varying based upon both the size of the muscle to be injected and the extent of muscle paralysis desired (i.e. amount of diopter correction desired);(6) to treat upper limb spasticity following stroke by intramuscular injections of BOTOX® into five different upper limb flexor muscles, as follows:                (a) flexor digitorum profundus: 7.5 U to 30 U        (b) flexor digitorum sublimis: 7.5 U to 30 U        (c) flexor carpi ulnaris: 10 U to 40 U        (d) flexor carpi radialis: 15 U to 60 U        (e) biceps brachii: 50 U to 200 U. Each of the five indicated muscles has been injected at the same treatment session, so that the patient receives from 90 U to 360 U of upper limb flexor muscle BOTOX® by intramuscular injection at each treatment session;(7) to treat migraine, pericranial injected (injected symmetrically into glabellar, frontalis and temporalis muscles) injection of 25 U of BOTOX® has showed significant benefit as a prophylactic treatment of migraine compared to vehicle as measured by decreased measures of migraine frequency, maximal severity, associated vomiting and acute medication use over the three month period following the 25 U injection.
Additionally, intramuscular botulinum toxin has been used in the treatment of tremor in patients with Parkinson's disease, although it has been reported that results have not been impressive. Marjama-Jyons, J., et al., Tremor—Predominant Parkinson's Disease, Drugs & Aging 16(4); 273-278:2000.
Treatment of certain gastrointestinal and smooth muscle disorders with a botulinum toxin are known. See e.g. U.S. Pat. Nos. 5,427,291 and 5,674,205 (Pasricha). Additionally, transurethral injection of a botulinum toxin into a bladder sphincter to treat a urination disorder is known (see e.g. Dykstra, D. D., et al, Treatment of detrusor-sphincter dyssynergia with botulinum A toxin: A double-blind study, Arch Phys Med Rehabil 1990 January; 71:24-6), as is injection of a botulinum toxin into the prostate to treat prostatic hyperplasia. See e.g. U.S. Pat. No. 6,365,164 (Schmidt).
U.S. Pat. No. 5,766,605 (Sanders) proposes the treatment of various autonomic disorders, such as hypersalivation and rhinitis, with a botulinum toxin.
Furthermore, various afflictions, such as hyperhidrosis and headache, treatable with a botulinum toxin are discussed in WO 95/17904 (PCT/US94/14717) (Aoki). EP 0 605 501 B1 (Graham) discusses treatment of cerebral palsy with a botulinum toxin and U.S. Pat. No. 6,063,768 (First) discusses treatment of neurogenic inflammation with a botulinum toxin. Erectile dysfunction has been reported as a symptom of botulism. Jenzer G., et al., Autonomic dysfunction in botulism B: a clinical report, Neurology 1975; 25:150-153; Naumann M. et al., Pure autonomic dysfunction in botulism type B, Naunyn Schmiedeberg's Archives of Pharmacology June 2002 (supp 2); 365 (abstract 89 at R31). This may be a result of circulating botulinum toxin present in a patient with botulism acting to block release of acetylcholine from cholinergic parasympathetic nerve endings in the corpora cavernosa of the penis. This would cause an inhibition of penile smooth muscle relaxation and therefore a reduced flow of blood into penile structures, and hence a flaccid penis. Contrarily, it has been speculated that a botulinum toxin can be used to cause an erection of the penis. Jones D. High performance. Nature 1989; 3:348.
It is known that botulinum toxin type A can have an efficacy for up to 12 months (European J Neurology 6 (Supp 4): S111-S1150:1999), and in some circumstances for as long as 27 months. The Laryngoscope 109:1344-1346:1999. However, the usual duration of an intramuscular injection of BOTOX® is typically about 3 to 4 months. The success of botulinum toxin type A to treat a variety of clinical conditions has led to interest in other botulinum toxin serotypes. Additionally, pure botulinum toxin has been used to treat humans. see e.g. Kohl A., et al., Comparison of the effect of botulinum toxin A BOTOX® with the highly-purified neurotoxin (NT 201) in the extensor digitorum brevis muscle test, Mov Disord 2000; 15 (Suppl 3):165. Hence, a pharmaceutical composition can be prepared using a pure botulinum toxin.
The botulinum toxin molecule (about 150 kDa), as well as the botulinum toxin complexes (about 300-900 kDa), such as the toxin type A complex are also extremely susceptible to denaturation due to surface denaturation, heat, and alkaline conditions. Inactivated toxin forms toxoid proteins which may be immunogenic. The resulting antibodies can render a patient refractory to toxin injection. A commercially available botulinum toxin containing pharmaceutical composition is sold under the trademark BOTOX® (available from Allergen, Inc., of Irvine, Calif.). BOTOX® consists of a purified botulinum toxin type A complex, albumin, and sodium chloride packaged in sterile, vacuum-dried form. The botulinum toxin type A is made from a culture of the Hall strain of Clostridium botulinum grown in a medium containing N—Z amine and yeast extract. The botulinum toxin type A complex is purified from the culture solution by a series of acid precipitations to a crystalline complex consisting of the active high molecular weight toxin protein and an associated hemagglutinin protein. The crystalline complex is re-dissolved in a solution containing saline and albumin and sterile filtered (0.2 microns) prior to vacuum-drying. BOTOX® can be reconstituted with sterile, non-preserved saline prior to intramuscular injection. Each vial of BOTOX® contains about 100 units (U) of Clostridium botulinum toxin type A complex, 0.5 milligrams of human serum albumin and 0.9 milligrams of sodium chloride in a sterile, vacuum-dried form without a preservative.
To reconstitute vacuum-dried BOTOX® sterile normal saline without a preservative; 0.9% Sodium Chloride injection is used by drawing up the proper amount of diluent in the appropriate size syringe. Since BOTOX® is denatured by bubbling or similar violent agitation, the diluent is gently injected into the vial. BOTOX® should be administered within four hours after reconstitution. During this time period, reconstituted BOTOX® is stored in a refrigerator (2° to 8° C.). Reconstituted BOTOX® is clear, colorless and free of particulate matter. The vacuum-dried product is stored in a freezer at or below −5° C. BOTOX® is administered within four hours after the vial is removed from the freezer and reconstituted. During these four hours, reconstituted BOTOX® can be stored in a refrigerator (2° to 8° C.).
Other commercially available botulinum toxin containing pharmaceutical compositions include DYSPORT® (Clostridium botulinum type A toxin haemagglutinin complex with albumin and lactose in the formulation, available from Ipsen Limited, Berkshire, U.K. as a powder to be reconstituted with 0.9% sodium chloride before use), and MYOBLOC™ (an injectable solution comprising botulinum toxin type B, human serum albumin, sodium succinate, and sodium chloride at about pH 5.6, available from Solstice Neurosciences, Inc).
What is needed therefore is a simple method for treating premature ejaculation and/or prolongation of climax time. In particular, a long lasting, non-systemic method for treating premature ejaculation and/or prolongation of climax time is desired that does not entail oral or repeated ingestion of a pharmaceutical compound prior to engaging in sexual activity.

Method for treating premature ejaculation with a botulinum neurotoxin


An assembly of a pallet component and fasteners makes a pallet on which various kinds of objects can be placed and which is movable by a forklift. The pallet component includes two outer unit members and an intermediate connector. Each outer unit member has an inner side formed with spaced projections and concavely provided with spaced receiving recesses adjacent thereto. The spaced projections of each outer unit member correspond in position to and are therefore receivable in the spaced receiving recesses of the other outer unit member. The intermediate connector is bilaterally provided with connecting sections that have different orientations. The working end of each connecting section has spaced projections and spaced receiving recesses adjacent thereto so as to connect with the corresponding outer unit member. Thus, a pallet can be easily made according to the widths of objects to be placed on the pallet.

1. Technical Field
The present invention relates to pallets capable of supporting various kinds of objects and movable by a forklift to desired locations. More particularly, the present invention relates to an assembly of a pallet component and fasteners that is applicable to making a pallet. The pallet component is composed of at least two structural elements, depending on the required dimension of the pallet component, so as to facilitate the making of a stable pallet in accordance with the widths of objects to be placed on the pallet. Thus, not only are the mold manufacturing costs and production costs of the pallet significantly reduced, but also a plurality of such pallet components can be stacked in a space-efficient way during transportation.
2. Description of Related Art
Conventional pallets for supporting various kinds of objects are typically made by cutting wood boards to the desired sizes and nailing the wood boards according to predetermined configurations. As wood pallets are costly, plastic injection-molded pallets have been developed for supporting finished goods and being transported by a forklift. The injection molding of plastic pallets requires molds that are manufactured in conformity with the specifications of the pallets, as well as a machine capable of accommodating the molds so that a plastic material can be injection-molded therein. Therefore, if pallets of relatively large sizes are to be made by injection molding, the mold manufacturing costs will be high, and the injection molding machine must be increased in size. Moreover, as pallets of different dimensions must be formed in different molds and different injection molding machines, the overall equipment costs will be considerable. Besides, if a factory is not spacious enough for installing injection molding machines designed for making large pallets, the factory is simply unsuitable for producing such large pallets. Furthermore, large pallets usually cannot be stacked in a space-efficient way and therefore take up much space.

Assembly of pallet component and fasteners


A paint stripper for use by immersion of a painted substrate in a bath of the composition or application in place has two phases, one aqueous and the other of partially water soluble organic solvent. The aqueous phase is saturated with organic solvent. The organic solvent is preferably benzyl alcohol, dibasic ester or ethyl-e-ethoxypropionate, The total amount of organic solvent in the bath is suitably in the range 2 to 20%. The stripper may be acid or alkali activated. The application stripper is thickened to avoid separation during storage.

This is a national stage application of PCT/GB97/02917 filed Oct. 22, 1997.
The present invention relates to a method of stripping paint from a painted substrate, especially a metal substrate. The stripper comprises an aqueous phase and an organic phase. A wide range of organic solvents have been used in paint stripper formulations. The most successful solvents are chlorinated hydrocarbons; especially methylene chloride. For environmental reasons, however, it is desirable for the levels of organic solvents, especially chlorinated solvents, to be reduced. The present invention allows reduced levels of chlorinated solvents to be used or even allows the use of such solvents to be avoided all together.
It is known to use water as an activator compound in organic solvent based paint strippers. Such compositions tend to have low levels of water, for instance less than 10%.
Other paint stripper compositions comprising both water and organic solvents which are immiscible with water have been described. Generally emulsifiers are added in quantities such that an emulsion of one phase in the other phase is formed. Such emulsions are storage stable and are required not to separate before use. Alternatively cosolvents may be added to create a homogeneous single phase system. Some examples are described for instance in DE-A-3325166, GB 1114220, EP-A-0611214 and US-A-4619706.
We have now surprisingly found that paint stripper compositions comprising immiscible (i.e. only partially soluble) organic solvent and water, but which are not in the form of an emulsion, can be used as paint strippers with very useful properties.
In a new method of stripping paint from a painted substrate surface according to the invention, the surface is contacted with the aqueous phase of a two-phase liquid stripper composition comprising an aqueous phase which comprises water, dissolved water-soluble activator compound and an organic solvent which has a solubility in water in the range 0.1 to 10% by weight at 20xc2x0 C. the organic solvent being present in the aqueous phase at a concentration of about the saturation level (as at the ambient conditions for the method) and comprising also an organic phase comprising the said organic solvent, which organic phase is in interfacial contact with the aqueous phase.
The inventors believe that it is the aqueous phase of the stripper composition which provides the primary stripping function, that is which attacks the paint on the substrate surface, thereby enabling its removal. It has been found that some thermal or mechanical agitation may enhance stripping. This enhancement may be due to provision of some solvent contact with the substrate. The solvent phase primarily provides a reservoir for dissolved solvent in the aqueous phase. In view of the continuous interfacial contact, as solvent is dragged out of the aqueous phase by contact with paint surfaces, it is replenished from the solvent phase. The solvent thereby remains dissolved in the aqueous phase at saturation level.
It is preferred for organic solvents to be used which have a solubility in water at 20xc2x0C. in the range 0.5 to 6.0% by weight. The level of dissolved solvent in the aqueous phase during the paint removal method may be higher than the end of that range, since it is possible for the methods to be carried out under conditions at which the solubility of the solvent in water is higher. Thus where the method is carried out at raised temperatures, the level of organic solvent in the aqueous phase may be at least 50% higher than the saturation level at 20xc2x0 C. for instance at least 100% higher or more.
The organic solvent may include chlorinated hydrocarbons, for instance in combination with other organic solvents. Thus the invention allows lower levels of chlorinated solvents to be used when a combination of such solvent and another organic solvent with which the chlorinated solvent is miscible is used. Preferably, however, the stripper composition is free of chlorinated organic solvents.
Suitable organic solvents are, for instance, benzyl alcohol, methylene chloride, dibasic ester, which is an oxygenated solvent composed of a mixture of methyldiesters of glutaric, succinic and adipic acids, or ethyl-3-ethoxy propionate. The solubility of these solvents in water are benzyl alcohol 3.3%, methylene chloride 1.32%, dibasic ester 5.6% and ethyl ethoxypropionate 2.9%.
The dissolved activator compound present in the aqueous phase of the composition used in the invention may be any compound which increases the paint stripper activity. Such compounds may be selected for the specific paint which is to be removed and/or the substrate from which the paint is to be removed, in order to provide optimum paint removal characteristics and/or minimise damage to the underlying substrate. The compound may often be a pH modifying compound. Such pH modifiers may be acids and considerable benefits have been observed with such acid activated systems. Acid activators are usually organic acids, for instance trichloroacetic, alkyl benzene sulphonic acid, benzoic or lactic acid. Preferably a pH modifying compound is an alkali. Suitable alkalis are, for instance, alkali metal hydroxides, alkaline earth metal hydroxides, silicates, for instance of alkali metals and organic basic compounds such as amines or ammonium compounds.
Another category of activator compound is an oxidising agent. Such oxidising agents are found to improve the performance of the aqueous phase. One example of an oxidising agent is hydrogen peroxide. Another example is permanganate, usually potassium permanganate.
The composition may, in addition to one of the above mentioned activator compounds, contain a secondary activator, such as an alcohol, for instance a C1-4 alcohol, preferably methanol.
The paint stripper composition may optionally contain surfactants, generally in low amounts such that the surfactant does not lead to the formation of an emulsion between the immiscible aqueous and organic phases. The inclusion of such surfactants may improve wetting of the substrate surface. Suitable surfactants are, for instance, alkyl benzene sulphonic acids, and fluorosurfactants. The surfactant is, generally, present in an amount of less than 2% by weight, more preferably less than 1% by weight, based on the total stripper composition weight.
Where the activator compound is a pH modifier, the pH is preferably above 9, more preferably above 10, or below 5, preferably below 4.
The present invention is of particular value where the stripping method uses an immersion technique, in which the article from which paint is to be stripped is dipped into a bath containing the stripper composition. In such embodiments, the stripper composition is generally present as a two-phase system with top and bottom continuous layers. Preferably the bottom layer is the organic solvent layer, that is the organic solvent preferably has a higher density than water. In this embodiment, the article from which paint is to be stripped is immersed in the top aqueous phase only and not into the bottom organic phase. After immersion for a suitable period of time, the article is removed from the bath.
The bath is reused for stripping paint from further articles. Where the bath is reused, it is generally necessary to replenish the bath to maintain an appropriate mixture of ingredients in the top aqueous phase. It is found that both solvent and water are dragged out of the bath whilst in addition the activator compound and solvent are consumed. Replenishment of the bath is straightforward in the invention which is convenient for the user. Thus since the bottom organic phase provides a reservoir of solvent for dissolution into the top aqueous phase, as the volume of the top phase becomes lower, it can be replenished merely by adding water. The aqueous phase maintains its saturation level of solvent since solvent can pass from the organic phase into the aqueous phase across the interfacial layer. The level of activator compound is maintained at the desired level by a continuous monitoring system and addition of activator compound either in neat form or in the form of a concentrated replenishment solution, usually in water. It is possible to use automated replenishment to control the total volume and activator compound concentration of the aqueous phase.
The total level of water in the stripper composition as a whole is generally in the range 50 to 99.5% by weight, preferably in the range 60 to 98% by weight, more preferably in the range 70 to 95%, especially 80 to 90% by weight.
An alternative embodiment of the stripping method of the invention, for non-immersion purposes, uses an application in place system, in which the stripper composition is generally brushed, sprayed or otherwise applied to the painted surface of a large substrate. Since such surfaces will include non-horizontal surfaces, in order to ensure that adequate levels of stripper composition are retained in contact with the painted substrate over sufficient periods of time, it is generally desirable to use a thickened composition. The thickened compositions may comprise a continuous aqueous phase and a dispersed organic phase. The organic solvent can cross the interface between the phases thereby providing a reservoir for solvent in the aqueous phase as in the first embodiment of the invention. The dispersion may be storage stable by virtue of the provision of a thickened continuous aqueous phase (as opposed to being an emulsion of dispersed organic phase in continuous aqueous phase).
Suitable thickeners for use in this embodiment of the invention must provide a thickening ability for the aqueous phase in the presence of the organic solvent at saturation levels and in the presence of the activator compound. Although naturally derived thickeners such as cellulose and starch derivatives may be used, the present inventors have found that improved performance and stability can be achieved using synthetic thickeners, for instance acrylic based associative thickeners such as are available under the trade mark Rheox or Rheovis. Inorganic thickeners, such as clays, for instance bentonite especially surface treated bentonite, may alternatively be used.
In the stripping method of the present invention, the substrate should, after having been in contact with the stripping composition for an adequate time to loosen the paint, be rinsed to remove stripper composition and loosened paint. Where the substrate has been immersed in a bath of the stripper composition, the article is removed from the bath and rinsed, for instance by immersion and/or spraying with water or an aqueous rinse solution. It may be desirable to use a pressurised water spray or to apply other mechanical assistance to remove loosened paint from the substrate. Where the stripper composition is applied in place, it is rinsed by a spray of water or aqueous rinse solution, for instance using a pressurised water spray.
The method of the present invention may be carried out under ambient, room temperature conditions. Alternatively the temperature of the stripper composition may be raised for instance where the composition is applied by immersion. Thus a bath of stripper composition may be heated to a temperature at which the stripping rate is improved. The present inventors have found that temperatures in the range 40 to 90xc2x0 C. can be used. Generally it is found that improved results can be achieved when the temperature is raised to above 50xc2x0 C. or even above 60xc2x0 C. although it is generally unnecessary to use temperatures of more than 80xc2x0 C. Thickened compositions are generally applied at ambient temperatures.
Where the composition is contacted with the substrate by immersion, it may be desirable for the body of stripper to be mechanically agitated. Such agitation should generally be inadequate to provide any stability to the two phase mixture against separation. Thus although such agitation may result in the temporary creation of a dispersed phase, this dispersed phase mixture will rapidly separate into two phases upon removal of the mechanical agitation.

Compositions and method for removing paint from a substrate


A novel composition and method for solubilizing amphiphilic drugs in a small amount of organic solvent for use in improved liposomes is disclosed. A phosphatidylglycerol is acidified in a small amount of organic solvent. The amphiphilic drug, such as Amphotericin B, suspended in organic solvent is then added to the acidified phosphatidylycerol and a soluble complex is formed between the phosphatidylglycerol and the amphiphilic drug. When the liposome composition incorporating the soluble complex is hydrated, the final pH of the hydrating aqueous buffer is carefully controlled. The Amphotericin B liposomes formed have markedly reduced toxicity.

This invention relates to a novel procedure for solubilizing amphiphilic drugs. In another aspect, this invention relates to improved methods of Amphotericin B liposome preparation. In another aspect this invention relates to improved methods of producing liposomes by a commercially feasible process. This invention also relates to liposomal Amphotericin B having reduced toxicity and to a new method of treatment with liposomal Amphotericin B.
Systemic fungal infections are a major cause of mortality in cancer patients and other immunocompromised individuals. Unfortunately, fungal infections very often defy treatment because the few drugs that destroy fungi are extremely toxic to the host. Because of the drugs"" toxicity, the lowest possible effective doses should be given. Unfortunately, because the drugs are diluted in the blood, and because large amounts of the drugs are degraded, or excreted or taken up by uninfected tissue, large doses actually are and must be given if the treatment is to be effective.
The preferred treatment for systemic fungal infections is primarily limited to two groups of drugs: the polyene antibiotics such as Amphotericin B and nystatin, and the imidazoles, such as ketaconazole and miconazole. The polyene antifungal antibiotics readily bind to sterol components of host cells causing disruption of the membrane, cell permeability and lysis. Amphotericin B has thus been associated with acute hemolytic crisis. Further, because it is particularly toxic to kidney tissue, it has been associated with irreversible renal damage and even kidney failure, at therapeutic dosage levels. Medoff, G., Kabayashi, G. (1980) Strategies in treatment of systemic fungal infections. New England Journal of Medicine 302: 145-55; Cohen, J. (1982) Antifungal chemotherapy. Lancet ii: 532-37; Graybill, J. R., Craven, P. C. (1983) Antifungal agents used in systemic mycosis: activity and therapeutic use. Drugs 25: 41-62.
It is a major goal of medical research to overcome the problems presented by the need for compromise between dosages high enough to control infection on the one hand, and unacceptable damage to healthy tissues on the other. Recently it has been discovered that needed doses of medicine can be delivered to diseased tissue while bypassing healthy tissue using certain liposomal formulations. Additionally, it has been recognized that medication can be incorporated into liposomes, microscopic delivery vesicles made, in part, of phospholipids. See U.S. Pat. No. 4,663,167xe2x80x94Composition and Method for Treatment of Disseminated Fungal Infections in Mammals, incorporated here by reference, and Vestar application Ser. No. 899,064 entitled xe2x80x9cImproved Treatment of Systemic Fungal Infections with Phospholipid Particles Encapsulating Polyene Antifungal Antibioticsxe2x80x9d (now U.S. Pat. No. 5,043,107), also incorporated by reference, . . . xe2x80x9dwhich discloses liposomal delivery vesicles made, in part, from phospholipids.
Phospholipids form closed, fluid filled spheres when mixed with water. Phospholipid molecules are polar, having a hydrophilic ionizable head, and a hydrophobic tail consisting of long fatty acid chains. Thus, when sufficient phospholipid molecules are present with water, the tails spontaneously herd together to exclude the water while the hydrophilic phosphate heads form bonds with the water.
The result is a bilayer in which the fatty acid tails point into the newly formed membrane""s interior and the polar heads point toward the aqueous medium. The polar heads at one surface of the membrane point toward the liposome""s aqueous interior and those at the other surface point toward the aqueous exterior environment. It is this chemical tendency to form liquid filled spheres that allows the liposome to be loaded with medication. As the liposomes form, water soluble molecules will be incorporated into the aqueous interior, and lipophilic molecules will tend to be incorporated into the lipid bilayer. Liposomes may be either multilamellar, like an onion with liquid separating many lipid bilayers, or unilamellar, with a single bilayer surrounding an entirely liquid center.
In studies of mice, Amphotericin B incorporated into liposomes has been shown to treat systemic fungal infections more effectively than when given as the free drug. Liposomes are not themselves toxic, and they protect their loads from being degraded or diluted. Thus, liposomes are thought to deliver concentrated doses of antifungal antibiotic at the diseased tissue without the toxicity that would otherwise be associated with freely circulating drug. Therefore, liposomal Amphotericin B drug doses can exceed the maximum tolerated dose of free Amphotericin B. Mehta, R. (1982), Liposomal aphotericin B is toxic to fungal cells but not to mammalian cells. Biochimica et Biophysica Acta 770: 230-34. Liposomal encapsulated Amphotericin B has also been shown to be an effective treatment for murine systemic fungal infections, including Candidiasis, Cryptococcosis, and Histoplasmosis. Graybill, J. R. et al (1983) Treatment of murine cryptococcosis with liposomal associated Amphotericin B. Journal of Infectious Diseases 145: 748-52; Taylor, R. L. et al (1982) Amphotericin B in liposomes: A novel therapy for histoplasmosis. American Review of Respiratory Diseases 125: 610-611.
Liposomal Amphotericin B has also shown effectiveness in human patients, life saving when other treatments have failed, including freely circulating Amphotericin B. Systemic fungal infections are seen most commonly in people whose immune systems are compromised by disease or immunosuppressive drug therapy. As previously mentioned, these infections are a common cause of death to victims of acquired immune deficiency syndrome and to cancer patients undergoing chemotherapy. The causative agents of these fungal infections are often endogenous fungi that would be rendered harmless but for the patient""s impaired resistance. Lopez-Berestein, G. et al (1987) Treatment of hepatosplenic Candidiasis with liposomal Amphotericin B. Journal of Clinical Oncology 5:310-17.
Unfortunately, because of the chemical properties of the polyene antifungal antibiotics, it has heretofore not been possible to produce liposomal Amphotericin B in commercial quantities. These antifungal agents are called polyenes because they contain three to seven conjugated double bonds in the aliphatic chain making up a large lactone ring. The double bonds are incorporated into one side of the ring of 26 to 44 carbon atoms and along the opposite side of the ring 6 to 12 hydroxyl groups are present. Additionally, these molecules contain specific carboxylic acid groups and amine groups. Amphotericin B and nystatin, for example, possess both an aminosugar and a carboxylic acid group. The polyene regions of the molecules are of course hydrophobic and lipophilic while the polyol and ionizable regions are hydrophilic and lipophobic. As such, these molecules are called amphiphilic. Additionally, because of the carboxylic group and the amine group, Amphotericin B can act as a Lowry-Brxcfx86nsted acid or proton donor, or as a Lowry-Brxcfx86nsted base or proton acceptor. The combination of these functionalities makes polyenes very poorly soluble in water and most organic solvents. Bennett, J. E. (1974) Chemotherapy of systemic mycoses. New England J. Medicine 290: 320-23.
It has been the persistent problem of insolubility of the polyene antifungal antibiotics in general and of Amphotericin B in particular that has previously limited the prior art. Liposomal Amphotericin B formation was heretofore limited to generally two methods, described below, neither feasible for commercial scale production and neither showing long-term stability or as great a reduction in toxicity as the preparations herein described.
One prior method requires that the Amphotericin B be first dissolved in large volumes of volatile organic solvent such as methanol. To that solution would then be added the lipid mixture dissolved in a volatile organic solvent such as methanol and/or chloroform. The solvents would then have to be removed from the mixture to form a lipid-Amphotericin B film. Removal of solvents could be accomplished by a variety of methods but usually by evaporation to dryness in a round bottom flask under vacuum or nitrogen. Taylor, R. L. (1980) Amphotericin B in liposomes: A novel therapy for histoplasmosis. Am. Review Respiratory Disease 125: 610-11; Graybill, J. R. et al (1982) Treatment of murine crytococcosis diseases. J. Infectious Diseases 145: 748-52; Lopez-Berestein, G. (1983) Treatment and prophylaxis in disseminated infection due to Candida albicans in mice with liposome-encapsulated Amphotericin B. J. Infectious Diseases 147: 939-45; U.S. Pat. No. 4,663,167xe2x80x94Composition and method for treatment of disseminated fungal infections in mammals. The prior art methods thus required removal of large volumes of organic solvent, and preparation of a lipid-Amphotericin B film, an extra step eliminated by the present invention. Moreover, the prior art methods were thus practicable only in discrete batches and were not amenable to the continuous flow process of the current invention. These are two disadvantages which the industry has long attempted to overcome.
In another method of forming liposomal Amphotericin B, the lipid mixture is dissolved in chloroform or another solvent and deposited and dried on the sides of a round bottom flask or vesicle surface. A solution of Amphotericin B dissolved in a small amount of dimethyl sulfoxide would then be added to the previously deposited lipid film. The resulting preparation would thereafter have to be dialyzed against a buffered saline or other solution to remove the dimethyl sulfoxide and non-intercalated Amphotericin B. The procedure was extremely time consuming and expensive, and typically resulted in incorporation of only 70% of the initial Amphotericin B. Tremblay, C. et al (1984) Efficacy of liposome-intercalated Amphotericin B in treatment of systemic Candidiasis in mice. Antimicrobial Agents and Chemotherapy 26: 170-73.
Prior to the present invention, it was not possible to dissolve Amphotericin B in small quantities of volatile solvent, such that scaled-up production could be commercially feasible. The present invention also enables the dissolved Amphotericin Bxe2x80x94phospholipid liposomal solution to be spray dried, thus making commercial quantities practical by elimination of the elaborate and time consuming steps detailed above.
Accordingly, one object of the present invention is to provide an improved process for the solubilization of amphophilic drugs.
Another object of the present invention is to provide an improved process for the encapsulation of polyene antifungal antibiotics into liposomes.
More specifically, an object of the present invention is to provide an improved process for the formation of liposomal Amphotericin B.
Yet another object of the present invention is to provide a commercially feasible process for the production of liposomal Amphotericin B.
Yet another object of the present invention is to provide a process for the formation of liposomal Amphotericin B with reduced toxicity.
A further object of the present invention is to provide new methods of treatment with Amphotericin B.
The manner in which these and other objects are realized by the present invention will be apparent from the summary and detailed description set forth below.
According to the present invention, the steps of either evaporation of the large volumes of volatile solvent into which the antibiotic is at least minimally soluble, or of removing a nonvolatile solvent in which the antibiotic and lipid has been dissolved, typically by dialysis, have been eliminated. Instead, the present invention provides a new and useful lipophilic charge complex of Amphotericin B which overcomes the previous problems of insolubility. Additionally, the present invention provides a new and useful process for the production of Amphotericin B liposomes suitable for scaling up production to commercial quantities. Additionally, the invention provides a method for production of Amphotericin B liposomes of increased stability and decreased toxicity.
In one preferred embodiment of this invention, the new soluble complex is formed between Amphotericin B and distearoyl-phosphatidylglycerol which has been protonated during dissolution in a solution of chloroform and methanol acidified to a pH of about 1.0 to 3.0. The Amphotericin Bxe2x80x94phospholipid complex, while in solution in the small amount of acidified chloroform and methanol, can be mixed with phosphatidylcholine and cholesterol and reproducibly spray dried under controlled conditions to yield a lipid powder which is readily processed into liposomes, using an aqueous buffer solution having a pH such that the pH of the final solution is below about 5.5, preferably between about 4.5 and 5.5. Accordingly, the present invention allows commercial scaling up of new materials for liposomal production. Further, the liposomes formed with this invention can be lyophilized and stored for later rehydration and injection without significant change in size or toxicity. The advantages of the present invention, will become clear after considering the following detailed description.
Distearoylphosphatidylglycerol, or other homologous phosphatidylglycerols, such as dilaurylphosphatidylglycerol, dimyristoylphosphatidylglycerol or others, is dissolved in an equal volume solution of chloroform and methanol and acidified before forming the described soluble complex with Amphotericin B. Thus, 2.5 N hydrochloric acid is added to the distearoylphosphatidylglycerol sodium salt solution in methanol:chloroform (1:1) to adjust the pH to between 1.0 and 3.0, as measured on prewetted pH paper, prior to mixing with Amphotericin B, and thereby acidifying the phospholipid. The Amphotericin B, or other polyene such as a tetraene, pentaene, or hexaene, is suspended in an equivolume solution of chloroform and methanol and then added to the acidic distearoylphosphatidylglycerol solution. Complex formation is facilitated by briefly warming the solution to about 65xc2x0 Centigrade. At this stage, the concentration of Amphotericin B may be in excess of 45 mg/ml.
In the resulting solution additional lipids such as phosphatidylcholines may also be dissolved. Cholesterol or another sterol, such as ergosterol, stigmosterol, or androsterone, is included to improve the stability of the resulting liposomes, and thus maintain the liposome intact during circulation in the bloodstream. The solution is a translucent orange. Typically, the Amphotericin B concentration in the final solution is greater than 25 mg/ml, and the total dissolved solid material 15-20 percent by weight.
Among the additional phosphatidylcholines that may be dissolved in the Amphotericin B, acidic phosphatidylglycerol complex solution, hydrogenated egg phosphatidycholine, hydrogenated soya lecithin and distearoyl or dipalmitoyl phosphatidylcholine are preferred such materials. Hydrogenated natural phospholipids or saturated aliphatic phospholipids are believed to work well because the lack of double bonds in the side chains is thought to render the liposomes resistant to oxidation and more physically stable.
The organic solvents may be removed from the solution by rotary evaporation in a round bottom flask leaving a dry film comprised of the complex and other lipid materials. Other equivalent methods of solvent removal are also suitable, such as drying under vacuum. Alternately, the solution can be applied to a spray dryer and solvent removed in a continuous process to produce large quantities of a free flowing yellow powder for liposome preparation. This novel complex thus provides the long awaited continuous production capability sought by industry.
After the last traces of organic solvent have been removed, the dried lipid complex powder may be stored for later use as a starting material for liposome preparation. This product affords the stability necessary for storage previously unavailable to the industry. Thus, the initial chemical steps need not be undertaken each time liposomes are desired.
Liposome preparation is accomplished by first hydrating appropriate quantities of the lipid complex powder with an aqueous buffer, preferably at about 65xc2x0 C. Aqueous buffer solutions may also contain salts such as sodium chloride, or saccharides such as dextrose or lactose, to achieve any desired osmolarity. The pH of the solution is carefully controlled to achieve a final solution having a pH of about 5.5 or less, generally between about 2.0 and 5.5, preferably between about 4.5 and 5.5.
Liposomes are then formed by the application of shearing force. Typically shearing force can be applied using sonification or homogenization, or by freezing and thawing, dialyzing away a detergent solution from lipids, or other known methods used to prepare liposomes. The size of the liposomes, as well as whether they are multilamellar or unilamellar, can be controlled using a variety of known techniques including the duration of sonication. See Gregoriadis, G. A. Simple Procedure to Preparing Liposomes Capable of High Encapsulation Efficiency Under Mild Conditions. Liposome Technology, Gregoriadis, G. A. (ed.) CRC Press: Boca Raton, Florida (1983). The present invention is adaptable, in particular, to scaling up production of the small unilamellar liposomes disclosed in pending Vestar, Inc. application Ser. No. 899,064xe2x80x94Improved Treatment of Systemic Fungal Infections with Phospholipid Particles Encapsulating Polyene Antifungal Antibiotics, incorporated here by reference. Such small liposomes can be sterilized by filtration since their diameter is less than 0.2 xcexcm. Virtually all of the initial Amphotericin B becomes associated with the liposome fraction when this technique is employed.
These liposome preparations, when formed in saccharide solutions such as 9% lactose, may be lyophilized in vials under suitable conditions to form a dried yellow cake or plug of material. At a later time water may be introduced into the vial to redissolve the solid cake and form a suspension of Amphotericin B liposomes suitable for injection. Lyophilization thus affords the clinician the significant advantage of increased convenience.
Amphotericin B, and other polyene antifungal antibiotics, are amphiphilic. One side of the macrocyclic compound is composed of a series of unsubstituted hydrocarbons with double bonds while the opposing side is substituted with hydroxyl groups. Thus, the molecules tend to exhibit polarity, one side lipophilic and hydrophobic, the other side lipophobic and hydrophilic.
Furthermore, Amphotericin B has one carboxyl group, a Lowry-Brxcfx86nsted acid, and one amine group, a Lowry-Brxcfx86nsted base. Therefore, in a neutral pH range of from 5 to 9 the carboxyl group tends to give up a proton while the amine group tends to accept that proton. The net result is that the molecule remains neutral and uncharged while at the same time having two ionized or charged groupsxe2x80x94a negatively charged carboxyl group and a positively charged amine group. In that same neutral pH range phospholipid molecules, such as distearoylphosphatidylglycerol are charged. They have an ionized phosphate group giving the molecule a negative charge. Equally, such phospholipid molecules are amphiphilic in that the long alphatic tails are hydrophobic and lipophilic while the ionizable phosphate head is, of course, hydrophilic and lipophobic.
When, however, the phospholipid molecule is solubilized in a protic solvent with a pH between about 1.0 and 3.0, the phospholipid molecule tends to accept a proton and thus form a comparatively neutral molecule. When Amphotericin B is then added to the above acidified solution, the proton of the phosphate group will tend to be transferred to the carboxyl group of the Amphotericin B. The result is that the Amphotericin B molecule will have a net positive charge. Concomitantly, the phospholipid""s phosphate group will give up a proton and become negatively charged. The thus formed, oppositely charged, molecules attract; their oppositely charged groups forming an ion pair.
The molecular attraction between Amphotericin B and phosphatidylglycerol is thus greatly increased. The aliphatic hydrocarbon chains of the phospholipids are attracted by hydrophobic interactions to the long chain of unsubstituted double bonds of the polyene. In the specific instance of Amphotericin B, the molecule is a heptaene with seven double bonded carbons along an unsubstituted section of 16 carbon atoms. In the specific instance of distearoylphosphatidylglycerol there are 16 unsubstituted methylene groups between the ester group and terminal methyl group.
In addition to the hydrophobic interaction, the ionized groups form a strong association. In the protonating environment above described, the Amphotericin B will have a positive charge and the phosphatidylglycerol will have a negative charge. Thus, the phosphatidylglycerol and the Amphotericin B will form a strong association.
The strongly associated complex so formed is highly soluble in small amounts of organic solvent. Thus, the disadvantages of the aforementioned prior art methods have been overcome. See, for example, U.S. Pat. No. 4,663,167, May 5, 1987, Lopez-Berestein et al. Accordingly, the present invention provides a procedure for scaling-up production which the industry has long awaited.
A further aspect of the present invention is that the Amphotericin Bxe2x80x94phosphatidylglycerol complex will associate with phosphatidylcholine and cholesterol during formulation and will not precipitate out of the organic solvent solution. Furthermore, if the pH of this organic solvent is maintained at 4.5 or less, the complex continues to remain stable and strongly associated. During liposome hydration, the pH of the aqueous buffer is controlled to give a solution having a final pH preferably between about 4.5 and 5.5. At that pH, the Amphotericin B-lipid complex is highly stable and has a high affinity for the lipid bilayer into which it becomes inserted. The result is a decrease in acute toxicity as demonstrated by the following Chart 1:
Thus, a significant decrease in toxicity is attained using the present invention. The decrease in toxicity allows for an increase in the therapeutic dosages that may be safely administered and thus offers greatly improved methods for Amphotericin B treatment. See U.S. Pat. No. 4,663,167, May 5, 1987, Lopez-Berestein et al.
Furthermore, the associated complex is highly stable during storage. The liposomes formed from either a film, or spray dried powder, after hydration with a saccharide buffer, may be lyophilized. The lyophilized cake can be stored preferably in a sterile lyophilization vial and later rehydrated with sterile water for injection. The reconstituted liposomes retain therapeutic efficacy.


A personal onboard information system for use in a vehicle. The system includes a portable, personal computer including a touch-responsive screen and a cradle mounted on the vehicle for detachably receiving the computer. The computer is programmed to generate characters of preset size on a display on the screen. The cradle has a lamp for illuminating the computer screen and a power supply for supplying the computer with power when the computer is in the cradle. The system further includes an interface communicating with the computer for formatting the display for use of the computer in the vehicle when the computer is in the cradle. The interface is resident in the computer or in the cradle and causes the display to display characters relatively larger than the characters of preset size so that the characters can be easily viewed by a passenger thereby facilitating use of the computer in the vehicle.

This invention generally relates to portable, personal computer systems and, in particular, such a system formatted for use in an automobile or other vehicle.
A portable, personal computer system which is small enough to be easily held in a user's hand offers numerous advantages over larger computer systems. Portability is one of the essential advantages of these palm-top, or hand-held, computers, often referred to as personal digital assistants (PDA). Unfortunately for persons traveling, though, a PDA is particularly ill-suited for use in a vehicle, such as an automobile.
In the environment of a vehicle, a PDA cannot be held in a user's hand while operating the vehicle. Among other reasons, using a PDA configured in the conventional manner while operating a vehicle could be dangerous and increase the risk of accident by unduly distracting the vehicle's operator and occupying the operator's hands. Thus, a means for supporting the PDA in the vehicle is required to free the hands of the PDA user for driving. However, such a support necessarily places the PDA further away from the user than holding the PDA in the user's hand. Due to the relatively small size of the PDA and its screen, the characters displayed on the screen are too small to be easily viewed unless the PDA is very near its user. Further, operation of the PDA is complicated because the user, whether operating the vehicle or merely a passenger in the vehicle, is unable to precisely select the desired key (or portion of the PDA screen) due to the small size of the characters and the movement of the vehicle. For these reasons, operation of a conventional PDA system in a moving vehicle can be difficult and very hazardous.
Further disadvantages of using conventional PDA systems in a vehicle, either by the vehicle's operator or a passenger, include the inability to view the PDA screen at night without turning on the vehicle's interior lights.
Also, the battery life of the PDA is limited and a power connection within the vehicle is desirable. Conventional PDA systems, however, are lacking because the ports and connectors for power, as well as serial access to the PDA's system board, are small and fragile. For this reason, these connectors are completely incompatible with use of the PDA in a moving vehicle.

Apparatus and method for a personal onboard information system


A portable computer has pivotally connected base and display screen lid housings and is provided with a generally wedge-shaped auxiliary component housing which is releasably latchable to the bottom of the base housing and extends across only a rear underside portion of the base housing. The attached auxiliary housing representatively carries a CD/DVD media drive unit and a floppy disk drive unit, operatively couples them to various computer components in the base housing, and is configured to rearwardly and upwardly tilt he base housing keyboard at a predetermined comfortable typing angle when the base housing is placed atop a horizontal work surface such as a desktop.

1. Field of the Invention
The present invention generally relates to electronic apparatus and, in a preferred embodiment thereof, more particularly relates to a portable notebook computer having removably secured to the bottom side of its base housing a specially configured auxiliary component housing.
2. Description of Related Art
Modern portable notebook computers are typically provided with a media storage device such as a floppy disk drive and/or a multimedia device such as a CD-ROM/DVD device. These media devices may be carried within the base housing portion of the notebook computer or be a separate portion of the overall portable computer system which must be externally connected to the base housing. In this case, the basic computer has a considerably smaller spatial envelope, and tends to be substantially lighter and more portable, due to a corresponding reduction in the required size of its base housing.
One conventional method of providing the computer with external media device capability is to simply provide, for example, an external floppy drive device which has a cable that may be releasably plugged into the computer base housing, with the externally floppy drive device sitting off to one side of the base housing on a work surface such as a desktop area. This method of selectively augmenting the functionality of a notebook computer tends to be rather cumbersome, and undesirably requires additional desktop work space.
Another previously proposed and somewhat more desirable technique for providing for the external connection to a notebook computer base housing of auxiliary computer components such as media devices has been to package such components in an additional rectangular housing which releasably attaches to the underside of the base housing and occupies its entire vertical footprint, thereby substantially increasing the height of the overall notebook computer base structure along the entire underside area of the base housing portion of the computer.
While this approach requires less workspace area when the computer (with the auxiliary housing operatively attached to the underside of its base housing) is placed atop, for example, a desktop, it presents various problems, limitations and disadvantages of its own. For example, as mentioned above, the entire height of the notebook computer is increased, thereby undesirably making the previously slim computer appear considerably thickerxe2x80x94a characteristic which may trigger buyer resistance in an era where notebook computers are increasingly being packaged in thinner and thinner packages.
Additionally, when the computer is placed on a horizontal work surface a conventionally configured additional housing attached to the underside of the computer base housing raises the computer keyboard to a potentially uncomfortable typing height while maintaining the elevated keyboard in a parallel relationship with the underlying desktop work surface. If the user prefers an upwardly and rearwardly sloping keyboard orientation, suitable support feet must be added to the overall computer apparatus to accommodate this user preference.
From the foregoing it can be readily seen that a need exists for an improved structure for packaging auxiliary components which may be removably attached to the underside of a portable computer base housing. It is to this need that the present invention is directed.
In carrying out principles of the present invention, in accordance with a preferred embodiment thereof, electronic apparatus is provided which is representatively in the form of a portable notebook computer having a base housing to which a display screen is pivotally secured. The base housing has a plurality of electronic components operatively disposed in its interior, and the apparatus further comprises an auxiliary housing having top and bottom sides and an interior in which at least one auxiliary component is operatively disposed. Illustratively, a media storage device, representatively a floppy disk drive, and a multimedia drive unit, representatively a CD ROM/DVD drive unit, are operatively disposed within the auxiliary housing.
The auxiliary housing is operatively securable to the bottom side of the base housing and is configured and positioned on the bottom base housing side in a manner such that when the base housing is placed atop a horizontal support surface the auxiliary housing causes the base housing (and thus the keyboard thereon) to be rearwardly and upwardly sloped relative to the support surface at a predetermined tilt angle which is representatively within the range of from about 8.5 degrees to about 9.5 degrees, and is preferably about 8.8 degrees.
According to features of the invention, the auxiliary housing has a generally wedge-shaped exterior spatial envelope, and extends along only a rear portion of the bottom side of the base housing when operatively attached thereto.
In the illustrated preferred embodiment of the electronic apparatus, the base and auxiliary housings have cooperatively interengageable electrical connectors thereon which, when interengaged, electrically couple the base and auxiliary housing components. The base and auxiliary housings also have cooperatively engageable latch structures which function, in response to placement of the auxiliary housing against the underside of the base housing, to automatically and releasably latch the auxiliary housing to the base housing. The auxiliary housing is preferably configured to be inset from opposite left and right ends of the base housing to reduce, from a visual aesthetic standpoint, the noticeable thickening of the base portion of the computer caused by the attachment thereto of the auxiliary component housing. To further expand the functionality of the electronic apparatus, a docking station electrical connector is disposed on the underside of the auxiliary housing to permit the computer to be placed atop and electrically coupled to an underlying computer docking station.

Portable computer with removable bottom component housing

Hewlett-Packard Development Company, L.P.


A novel boot method for a system, whose system BIOS has been disabled by erroneous microcode input as the result of inadvertent miscoding or malicious intent. The keyboard BIOS of the system is utilized in the present invention. If the CPU of the computer crashes during a microcode update, the system notes the error (setting a number of flags) and restarts the computer. The restarted system will not perform the microcode update, but goes straight to the normal BIOS activation, avoiding the crash problem. To fix the problem, users need only request the correct microcode from CPU retailers to amend the errors in the BIOS.

1. Field of the Invention
The present invention relates in general to a boot method for systems. In particular, the present invention relates to a boot method for a system whose system BIOS has been disabled by erroneous microcode input as the result of inadvertent miscoding or malicious intent.
2. Description of the Related Art
In the environment of the present-day microprocessor industry, life cycles of CPU products are dramatically shortened; CPU retailers, in order to react to short RandD cycles, publish CPU products first to reach the market, then update microcode to system manufacturers later. The microcode is rectified when posting BIOS systems to ensure the liability of the CPU products. Additionally, most CPU retailers require system manufacturers to provide an interface in the BIOS system, so after purchasing, users can enter the microcode provided by the CPU retailers into the BIOS system as needed. Thereby, each time the users start up, system BIOS updates the microcode to effect BIOS activation of the system, and posts the latest program edition into the system.
However, this method can easily be used by malicious parties as a way to damage systems. Although CPU retailers have adopted the checksum method to verify the microcode written in BIOS, the malicious parties can still fabricate the checksum codes or send virus messages to post the wrong microcode into the system, disabling the CPU.
An object of the present invention is to provide a novel boot method for a computer system. When a system BIOS code (or a microcode to be updated) in the system is damaged, the system is still able to complete BIOS activation procedure and perform the startup.
The keyboard BIOS is incorporated in the present invention. If the computer CPU crashes during the microcode update, the system records the event (setting a number of flags) and restarts the computer. The restarted system will not perform the update, but go straight to normal BIOS activation for the system, thus avoiding the crash problem. Users need only ask for the correct microcode from the CPU retailer to fix the errors in the BIOS.
To solve the described problems, the present invention provides a boot method for a system; the boot method allows the system to be rebooted in the event of erroneous microcode input; the boot method comprises the following steps: after activating the system, checks to determine the necessity for an update, and notification to the keyboard BIOS about the update; the keyboard BIOS waits for a first predetermined period after the update; if an updated signal is not received, the keyboard BIOS sets a fail flag, shuts down the system and, after a second predetermined period, reboots. If the fail flag has already been set during the previous boot, the system does not update the microcode after activation, and sets a non-update flag to allow the system to perform normal BIOS activation. If the non-update flag has already been set during the previous boot, the system starts the BIOS activation procedure after the computer is activated; otherwise, the system confirms whether the update should commence according to the fail flag settings.

Method for updating microcode in a system wherein a keyboard BIOS expects an updated signal within a time period after microcode update commences


A jar or bottle container for dispensing food items or other products that may have a liquid component and a solid component has a platform that is supported on a threaded post on the interior of the jar. A base seated at an open bottom of the jar body is turned to rotate the post and elevate the platform to push the food item upward. The platform has a ring seal and radial lip or tooth structure that rides in vertical channels formed in the inner wall of the jar body. A weight or ballast may be present in the base.

This invention is directed to a food product container that is especially suited for food products that are viscous mixtures of solid and liquid components, such as mayonnaise, peanut butter, jam, sauces, for example, and is more particularly directed to a jar that includes a platform that can be made to rise to push the product up towards the mouth of the jar for easy dispensing.
The invention is more specifically concerned with an Easy Glide Jar that is designed to allow the user to push the jar contents up to the top of the jar as the contents are consumed, both for convenience and to avoid waste.
Peanut butter, jam, sauces, mayonnaise, and other semi-liquid food products are typically stored in glass or clear plastic containers (i.e., jars) with twist off lids. As the food product is consumed, the remaining product resides at the bottom of the jar, and it is often difficult to reach in with a spoon or knife and remove that product. Accordingly, some means has been needed to push the contents up towards the mouth of the jar, but no suitable push-up system has been proposed for such viscous semi-liquid food products. There have been twist dispensers for various products, such as deodorant or other personal health products (See U.S. Pat. Nos. 7,270,495; 7,207,739; 6,918,511; and 5,137,185). These dispenser have used a threaded post that is rotated to move a cup so as to move a material, that is in stick form, to an open end of a tube. These dispenser containers are not adapted for viscous semi-liquids in the nature of jams, sauces, butters, or other products that have a liquid component that can settle out, and which would easily leak or drip past the elevator cup portion of the device. An example of a dispenser for shortening is shown in U.S. Pat. No. 2,656,953. That dispenser employs a rotary threaded post to drive a disk member down and urge the shortening through a dispensing tube. None of these previously proposed dispensing containers are particularly well suited for storing and dispensing peanut butter, sauce, jam, or any food product of that nature that comprises a liquid component and a solid component.