Methods and compositions are disclosed utilizing the optically pure (xe2x88x92) isomer of amlodipine. This compound is a potent drug for the treatment of hypertension while avoiding the concomitant liability of adverse effects associated with the racemic mixture of amlodipine. The (xe2x88x92) isomer of amlodipine is also useful for the treatment of angina and such other conditions as may be related to the activity of (xe2x88x92) amlodipine as a calcium channel antagonist such as cerebral ischemia, cerebral disorders, arrhythmias, cardiac hypertrophy, coronary vasospasm, myocardial infarction, renal impairment and acute renal failure, without the concomitant liability of adverse effects associated with the racemic mixture of amlodipine.

This invention relates to novel compositions of matter containing optically pure (xe2x88x92) amlodipine. These compositions possess potent activity in treating both systolic and diastolic hypertension while avoiding adverse effects including but not limited to edema of the extremities, headache and dizziness, which are associated with administration of the racemic mixture of amlodipine. Additionally, these novel compositions of matter containing optically pure (xe2x88x92) amlodipine are useful in treating angina and such other conditions as may be related to the activity of (xe2x88x92) amlodipine as a calcium channel antagonist including but not limited to cerebral ischemia, cerebral disorders, arrhythmias, cardiac hypertrophy, coronary vasospasm, myocardial infarction, renal impairment and acute renal failurexe2x80x94while avoiding the adverse effects associated with administration of the racemic mixture of amlodipine. Also disclosed are methods for treating the above-described conditions in a human while avoiding the adverse effects that are associated with the racemic mixture of amlodipine, by administering the (xe2x88x92) isomer of amlodipine to said human.
Many organic compounds exist in optically active forms, i.e., they have the ability to rotate the plane of plane-polarized light. In describing an optically active compound, the prefixes D and L or R and S are used to denote the absolute configuration of the molecule about its chiral center(s). The prefixes d and l or (+) and (xe2x88x92) are employed to designate the sign of rotation of plane-polarized light by the compound, with (xe2x88x92) or 1 meaning that the compound is levorotatory. A compound prefixed with (+) or d is dextrorotatory. For a given chemical structure, these compounds, called stereoisomers, are identical except that they are mirror images of one another. A specific stereoisomer may also be referred to as an enantiomer, and a mixture of such isomers is often called an enantiomeric or racemic mixture.
Stereochemical purity is of importance in the field of pharmaceuticals, where 12 of the 20 most prescribed drugs exhibit chirality. A case in point is provided by the L-form of the xcex2-adrenergic blocking agent, propranolol, which is known to be 100 times more potent than the D-enantiomer.
Furthermore, optical purity is important since certain isomers may actually be deleterious rather than simply inert. For example, it has been suggested that the D-enantiomer of thalidomide was a safe and effective sedative when prescribed for the control of morning sickness during pregnancy, while the corresponding L-enantiomer has been thought to be a potent teratogen.
The active compound of this composition and method is an optical isomer of the compound amlodipine, which is described in Campbell et al., U.S. Pat. No. 4,572,909. Chemically, this (xe2x88x92) isomer is 3-ethyl 5-methyl (xe2x88x92)-2-4-(2-chloropheny)-1, 4-dihydro-6-methyl- 3, 5-pyrinde dicarboxylate. The isomer will hereinafter be referred to as (xe2x88x92) amlodipine. (xe2x88x92) amlodipine also includes the substantially optically pure (xe2x88x92) amlodipine isomer.
Amlodipine, which is the subject of the present invention, is available commercially only as the 1:1 racemic mixture. That is, it is available only as the 1:1 mixture of optical isomers, called enantiomers. The racemic mixture of amlodipine that is commercially available for administration is a besylate salt.
The racemic mixture of amlodipine is in the class of compounds known as calcium antagonists. The concept of a specific mechanism of pharmacologic action related to the antagonism of calcium movement in the process of excitation-contraction was suggested by Fleckenstein et. al. (see Fleckenstein, A., Calcium Antagonism in Heart and Smooth Muscle: Experimental Facts and Therapeutic Prospects, New York, Wiley, 1983; Swamy, V. and D. Triggle, Modern Pharmacology, 2nd. Ed., Craig and Stitzel, Eds., Little, Brown and Co., Boston, 1986, Chapt. 26, 373-380; Triggle, D. J., and R. A. Janis, Ann. Rev. Pharm. and Tox. 27: 347-369, 1987). Many of the currently available calcium antagonists appear to antagonize the entry of calcium through voltage dependent channels in the plasma membrane of cells. The pharmacologic class of calcium antagonists consists of chemically diverse compounds. Given the structural heterogeneity of the class it is likely that the pharmacological action involves more than one site or mechanism of action.
Amlodipine is one of a series of dihydropyridine calcium antagonists. However, amlodipine has a generally slower onset and longer duration of action than, for example, nifedipine. (Jensen, H. et al., J. Hum. Hypertens., 42(5): 541-45, 1990). The metabolites of amlodipine apparently do not possess significant calcium channel blocking activity, while the parent drug offers a biological half-life of some 35-40 hours, prompting a once-daily dosage regimen. (Lorimer, A. R., et al., J. Hum. Hypertens., 3(3): 191-96, 1989; Glasser, S. F. et al., AJH, 2(3): 154-57, 1989). Its ability to block calcium channels in smooth muscle produces peripheral vasodilation resulting in decreases in both systolic and diastolic blood pressure.
With regard to the enantiomers of amlodipine, these are disclosed in Arrowsmith et al. J. Med. Chem., 29: 1696-1702 (1986). This reference discusses in vitro tests to determine calcium antagonist activity against calcium-induced constriction of potassium-depolarized rat aorta. The authors allege that in vitro evaluation of the enantiomers of amlodipine shows the (xe2x88x92) isomer to be twice as active as the racemic mixture in antagonizing calcium-induced constriction of potassium-depolarized rat aorta. The authors also allege that the (+) isomer of amlodipine is some 1,000 times less active in these in vitro tests. Further, European Patent Application No. 0 331 315 discloses a method for separation of the R(xe2x88x92) and S(+) isomers of amlodipine. While these references discuss the enantiomers of amlodipine, they only provide a method of synthesis and in vitro test results of the individual enantiomer activity as found in rat aorta. There is no disclosure of the effects of or a suggestion for administering one of the enantiomers of amlodipine to a human. Moreover, there is no disclosure or suggestion of the alleviation or reduction of side effects which is achieved by the administration of (xe2x88x92) amlodipine.
The racemic mixture of amlodipine is presently used primarily as an antihypertensive agent; it is generally taken orally as a once-daily therapy. As stated above, the racemic mixture of amlodipine produces peripheral vasodilation, resulting in decreases in both systolic and diastolic blood pressure when used as an antihypertensive agent. This antihypertensive effect occurs in the relative absence of significant or sustained effects on cardiac rate.
While yet the subject of extensive research, hypertension appears to be the product of an inherited predispositionxe2x80x94coupled with dietary, emotional, and environmental factors, which results in a structural adaptation of the cardiac muscle and the large blood vessels. Most patients display heightened vascular and cardiac reactions to sympathetic nervous stimulation, but the precise relationship of sympathetic nervous stimulation to the etiology of the disease. Nevertheless, hypertension results in chronic readjustment of cardiovascular hemodynamics, alteration of blood vessel walls, cardiovascular resistance and regional transmural pressures.
Pharmacologic management of hypertension is generally directed to the normalization of altered hemodynamic parameters, and many drugs and drug classes, either as monotherapy or in combination treatment, can reduce and control elevated blood pressure. However, treatment of hypertension does not always correspondingly benefit the morbidity and mortality of the condition, either because chronic hypertension has produced other significant and irreversible cardiovascular changes, or because present drugs have an adverse effect on some other risk factor for cardiovascular disease. Rather, current drug therapy simply provides sustained arterial pressure reduction.
Furthermore, the racemic mixture of amlodipine is useful in treating other disorders such as angina pectoris.
Angina pectoris is a highly variable, rather poorly understood clinical syndrome reflecting a myocardial ischemia. When cardiac work or myocardial oxygen demand exceeds the ability of the coronary arterial vascular system to supply oxygen, the resulting ischemia stimulates the sensory nerves of the heart, producing the sensation of angina characterized by episodes of precordial pressure, discomfort, or a severe, intense crushing pain which may radiate to several sites including the left shoulder and left arm. Physical activity or exertion characteristically initiates the condition, and rest or drug therapy relieves the condition. The signs and symptoms of an episode persist for a few minutes, but can be induced or exaggerated by a meal or exposure to cold air. Treatment is directed to the underlying disease, usually atherosclerosis, or to drugs which either reduce myocardial oxygen demand or improve oxygen supply. Calcium antagonists such as amlodipine have been particularly useful in treating vasospastic angina, the angina of effort, and the unstable angina, due to the effect of the calcium channel antagonist on cardiac and vascular smooth muscle.
Amlodipine may be useful in the treatment of cerebral ischemia. Cerebral ischemia, often the result of atherosclerotic disease or hypertension, results from insufficient cerebral circulation. Under normal circumstances, an extensive collateral circulation ensures adequate blood flow. However, cerebral ischemia may result from either an intra- or extracranial interruption of arterial blood flow. If interruption is transient, the cerebral tissues recover, and neurologic symptoms disappear. If the ischemia lasts for a somewhat more extended period, infarction results and the resulting neurologic damage is permanent. In the case of extended ischemia resulting in infarction, treatment is directed to the underlying vascular disease, to blood platelet aggregation inhibitors, and anticoagulant therapy.
Because of its activity as a calcium channel antagonist, amlodipine may also be useful in treating cardiac arrhythmias. Cardiac arrhythmias represent a broad, complex group of electrophysiologic disorders that affect the mechanical properties of the heart and vasculature, altering normal cardiac rhythm, function and output. Normal cardiac rhythm originates with the sinoatrial node, which possesses high intrinsic automaticity. Adequate automaticity and conduction lead to activation of atrial and ventricular fibers, producing in sequence the elements of normal functional heart beat. Calcium antagonists may be of value in conditions where calcium-related changes in membrane potential and conduction alter normal rhythm. In the absence of treatment, symptoms vary with individual arrhythmias, but are often the consequence of inadequate cardiac filling and output and often include fatigue, decreased exercise tolerance, syncope, shortness of breath, nausea, lightheadedness and the like.
Amlodipine may be useful to treat cardiac hypertrophy. Cardiac hypertrophy can result from excessive workload either due to an obstruction to outflow, termed systolic overload, or to excessive volumes presented to the heart in diastole, termed diastolic overload. Systolic overload results in concentric ventricular hypertrophy, in which there is an increased thickness in the walls of the heart not associated with increased volume. Diastolic overload causes dilation and hypertrophy with an increased blood volume. An inadequate cardiac output results from the heart""s failure in systolic or diastolic overload, leading to fatigue, shortness of breath, pulmonary congestion, edema and the like. Calcium channel antagonists effect workload and, as such, may be useful in treating cardiac hypertrophy due to the effect of the calcium antagonist on cardiac and vascular smooth muscle in reducing blood pressure.
It is also possible that amlodipine could be used to treat coronary arterial spasm. Coronary arterial spasm can occur in the absence of significant coronary atherosclerosis and is thought to be an initiating event in variant angina and in myocardial infarction. Coronary spasm may occur without the patient feeling any significant discomfort. In an electrically unstable heart, diverse neural impulses to the heart may provoke coronary vascular spasm. This may result in enhanced myocardial ischemia and arrhythmia, which in turn may culminate in ventricular fibrillation and sudden cardiac death. As in variant or vasospastic angina, the calcium channel antagonists may be of particular usefulness due to their effect on cardiac and vascular smooth muscle.
Furthermore, amlodipine may be useful in the treatment of myocardial infarction, ischemic myocardial necrosis, and ischemia reperfusion injury. Myocardial infarction or ischemic myocardial necrosis generally results from the abrupt reduction of coronary blood flow to a portion of the myocardium. The condition likely originates from atherosclerosis of the coronary arteries. Either coronary artery vasospasm or acute coronary thrombosis precipitates the condition, although the etiology is the subject of continuing research. Myocardial infarction is predominantly a disease of the left ventricle. Precordial pain and left ventricular dysfunction characterize the disease. The pain, which can be severe aching or pressure, leads to apprehension. Symptoms include left ventricular heart failure, pulmonary edema, shock or significant cardiac arrhythmia. Calcium channel antagonists may find utility in the management of myocardial infarction patients due to their effects on coronary artery vasospasm, blood pressure or other effects on cardiac function or vascular smooth muscle.
Amlodipine may be used to treat congestive heart failure. Congestive heart failure can be caused by hypertension, cardiomyopathy, coronary artery disease or valvular heart disease. Congestive failure results in poor cardiac output and elevated left-ventricular diastolic pressure, leading to dyspnea, fatigue, peripheral edema, and coughing. The ability of some calcium antagonists to lower afterload by dilating peripheral arteries without having a significant inotropic effect may increase their use in treating congestive heart failure.
Amlodipine may be of use in treating migraine. Classic migraine typically begins with visual auras followed by severe headaches, often accompanied by nausea and vomiting. Common migraine has similar symptoms without the preceding visual aura. The causes of migraine have been studied intensely, and are still a matter of debate. The most generally accepted cause is hypoxia due to reduced cerebral blood flow. Calcium channel antagonists have been used for migraine prophylaxis since they can increase cerebral blood flow.
Amlodipine may also be useful for treating Raynaud""s phenomenon, which is characterized by vascular spasm of the extremities. These vasospasms can be caused by cold or stress. A pallor or cyanosis is usually present due to severe constriction of the digital arteries. The phenomenon is often seen as a secondary disorder with arterial diseases or connective tissue diseases such as scleroderma, arthritis or lupus erythematosus. Calcium channel antagonists have been shown to be effective in treating Raynaud""s phenomenon.
Amlodipine may be useful in the treatment of asthma and bronchospasm. Symptoms of asthmaxe2x80x94coughing, wheezing, and dyspneaxe2x80x94are caused by constriction of tracheobronchial smooth muscle. Asthma attacks can be triggered by antigenic stimuli (pollen, dust) or non-antigenic stimuli (exercise, pollution, infection). The response to these stimuli lead to secretions of chemical mediators that cause smooth muscle contraction. Calcium channel antagonists can be used to control bronchoconstriction and relieve asthma attacks.
In addition, the racemic mixture of amlodipine may be useful to treat renal impairment and acute renal failure. Renal impairment and acute renal failure are clinical conditions of diverse etiology, which are associated with an increasing azotemia or urea nitrogen in the blood, and often an oliguria or a diminished volume of urine in relation to fluid intake. The pathophysiology may originate prerenally, manifest as inadequate renal perfusion, due to extracellular fluid volume depletion or cardiac failure. The most common cause of intrinsic renal failure is prolonged renal ischemia. Postrenal azotemia may be associated with obstruction or renal glomerular and tubular dysfunction. Laboratory findings disclose progressive azotemia, acidosis, hyperkalemia, and hyponatremia. Factors aggravating kidney impairment or failure must be specifically treated, including heart failure, obstruction and the like. Moderate or severe hypertension has a deleterious effect on renal function, and management of the hypertension with a variety of drugs including calcium channel antagonists may be useful therapy.
In addition, the racemic mixture of amlodipine could be useful in the treatment of cognitive disorders. Cognitive disorders include but are not limited to dementia and age-associated memory impairment.
Dementia can occur at any age. It is a structurally caused permanent or progressive decline in several dimensions of intellectual function that interferes substantially with individual normal social or economic activity.
One particular type of dementia is Alzheimers-type dementia. Alzheimers-type of dementia is thought to be due to a degenerative process, with a large loss of cells from the cerebral cortex and other brain areas. Acetylcholine-transmitting neurons and their target nerve cells are particularly affected. The brain shows marked atrophy with wide sulci and dilated ventricles. Senile plaques and neurofibrillary tangles are present. Memory loss is the most prominent early symptom. Disturbances of arousal do not occur early in the course. Alzheimer""s presenile and senile onset dementias are similar in both clinical and pathologic features, with the former commonly beginning in the 5th and 6th decades and the latter in the 7th and 8th decades. The dementia usually progresses steadily, becoming well advanced in 2 to 3 years. Some cases of dementia occurring in the presenile period are hard to classify and are sometimes labelled idiopathic or simple presenile dementia.
The signs and symptoms of dementia in particular Alzheimers-type dementia include depression, paranoia, anxiety or any of several other psychologic symptoms. The most common clinical picture is slow disintegration of personality and intellect due to impaired insight and judgment and loss of affect. Memory impairment increases, beginning with problems recalling recent events or finding names. The impairment varies greatly from time to time and often from moment to moment. Dementia generally is an insidious, slowly progressive, untreatable condition. However, the rate of progression varies widely and depends on the cause.
Another type of cognitive disorder is age-associated memory impairment (AAMI). AAMI is used to describe healthy non-demented people who have experienced memory loss over the course of the person""s life. Most commonly it is used to describe adults over the age of 50 who have experienced memory loss over the course of adult life. It has been estimated that between 25% and 50% of people over the age of 65 have this disorder.
Many calcium channel antagonists cause significant adverse effects. These adverse effects include but are not limited to tachycardia, orthostatic hypotension and fluid retention. In contrast to the situation with several other calcium channel antagonists, however, the racemic mixture of amlodipine has not been found to cause either marked or prolonged direct effects on heart rate or the reflex consequence of vasodilation.
However, the administration of the racemic mixture of amlodipine to a human has been found to cause still other adverse effects. These adverse effects include but are not limited to edema of the extremities including peripheral edema, headache, flushing/hot flashes, fatigue, vertigo, muscle cramps and dizziness.
Thus, it would be particularly desirable to find a compound with the advantages of the racemic mixture of amlodipine which would not have the aforementioned disadvantages of significant adverse side effects and which was useful for treatment of other conditions.
It has now been discovered that the optically pure (xe2x88x92) isomer of amlodipine is an effective antihypertensive agent for both systolic and diastolic hypertension, particularly in mild to moderate disease and angina, while avoiding adverse effects including but not limited to edema of the extremities, headache and dizziness, which are associated with the administration of the racemic mixture of amlodipine. It has also been discovered that these novel compositions of matter containing optically pure (xe2x88x92) amlodipine are useful in treating other conditions as may be related to the activity of (xe2x88x92) amlodipine as a calcium channel antagonists including but not limited to cerebral ischemia, cerebral disorders, arrhythmias, cardiac hypertrophy, coronary vasospasm, myocardial infarction, renal impairment and acute renal failure while avoiding the above-described adverse effects associated with the administration of the racemic mixture of amlodipine. The present invention also includes methods for treating the above-described conditions in a human while avoiding the adverse effects that are associated with the racemic mixture of amlodipine by administering the (xe2x88x92) isomer of amlodipine to said human.
The present invention encompasses a method of eliciting an antihypertensive effect in a human, while avoiding the concomitant liability of adverse effects, which comprises administering to said human in need of such antihypertensive therapy, an amount of (xe2x88x92) amlodipine or a pharmaceutically acceptable salt thereof, substantially free of its (+) stereoisomer, said amount being sufficient to alleviate hypertension, but insufficient to cause said adverse effects associated with administration of racemic amlodipine.
The present invention also encompasses an antihypertensive composition for the treatment of a human in need of antihypertensive therapy, which comprises an amount of (xe2x88x92) amlodipine or a pharmaceutically acceptable salt thereof, substantially free of its (+) stereoisomer, said amount being sufficient to alleviate said hypertension but insufficient to cause adverse effects of racemic amlodipine.
The present invention further encompasses a method of treating angina in a human, while avoiding the concomitant liability of adverse effects, which comprises administering to said human in need of such anti-angina therapy, an amount of (xe2x88x92) amlodipine, or a pharmaceutically acceptable salt thereof, substantially free of its (+) stereoisomer, said amount being sufficient to alleviate said condition but insufficient to cause said adverse effects associated with administration of racemic amlodipine.
In addition, the present invention encompasses an antianginal composition for the treatment of a human having angina, which comprises an amount of (xe2x88x92) amlodipine or a pharmaceutically acceptable salt thereof, substantially free of its (+) stereoisomer, said amount being sufficient to alleviate said angina but insufficient to cause adverse effects of racemic amlodipine.
A further aspect of the present invention includes a method of treating a condition caused by excessive calcium influx in cells in a human, while avoiding the concomitant liability of adverse effects, which comprises administering to said human in need of a reduction in excessive calcium influx an amount of (xe2x88x92) amlodipine, or a pharmaceutically acceptable salt thereof, substantially free of its (+) stereoisomer, sufficient to alleviate said condition but insufficient to cause said adverse effects of racemic amlodipine. Conditions caused by excessive calcium influx in cells in a human include but are not limited to cerebral ischemia, cerebral disorders such as cognitive disorders including but not limited to Alzheimer""s dementia and memory impairment, arrhythmias, cardiac hypertrophy, congestive heart failure, coronary vasospasm, migraine, bronchospasm and asthma, Raynaud""s phenomenon, myocardial infarction, renal impairment and acute renal failure.
Furthermore, the present invention includes a composition for treating a condition caused by excessive calcium influx in cells in a human, which comprises an amount of (xe2x88x92) amlodipine, or a pharmaceutically acceptable salt thereof, substantially free of its (+) stereoisomer, said amount being sufficient to alleviate said condition but insufficient to cause adverse effects of racemic amlodipine.
The commercially available racemic mixture of amlodipine (e.g., a 1:1 racemic mixture of the two enantiomers) causes antihypertensive and antianginal activity; however, this racemic mixture, while offering the expectation of efficacy, causes adverse effects. Utilizing the (xe2x88x92) isomer of amlodipine results in clearer dose-related definitions of efficacy, surprisingly diminished adverse effects, and accordingly, an improved therapeutic index. It is, therefore, more desirable to use the (xe2x88x92) isomer of amlodipine.
The term xe2x80x9cadverse effectsxe2x80x9d includes, but is not limited to, cardiovascular effects (including tachycardia and diminished contractility of the heart), edema of the extremities, headache, dizziness, flushing, fatigue, vertigo, and muscle cramps.
The term xe2x80x9csubstantially free of its (+) stereoisomerxe2x80x9d as used herein means that the composition contains a greater proportion or percentage of the (xe2x88x92) isomer of amlodipine in relation to the (+) isomer of amlodipine, said percentage being based on the total amount of amlodipine. In a preferred embodiment the term xe2x80x9csubstantially free of its (+) stereoisomer means that the composition contains at least 90% by weight of (xe2x88x92) amlodipine, and 10% by weight or less of (+) amlodipine. In the most preferred embodiment the term xe2x80x9csubstantially free of the (+) stereoisomerxe2x80x9d means that the composition contains at least 99% by weight (xe2x88x92) amlodipine, and 1% or less of (+) amlodipine. In another preferred embodiment the term xe2x80x9csubstantially free of its (+) stereoisomerxe2x80x9d as used herein means that the composition contains 100% by weight of (xe2x88x92) amlodipine. The terms xe2x80x9csubstantially optically pure (xe2x88x92) isomer of amlodipinexe2x80x9d and xe2x80x9coptically pure (xe2x88x92) isomer of amlodipine are also encompassed by the above-described meanings.
The term xe2x80x9celiciting an antihypertensive effectxe2x80x9d as used herein means providing a normalization to otherwise elevated systolic and/or diastolic blood pressure, and by so doing providing relief from any possible symptoms or other hemodynamic effects caused by the elevated pressure.
The term xe2x80x9ca method of treating anginaxe2x80x9d as used herein means relief from the symptoms of myocardial ischemia, which include, but are not limited to, episodes of precordial pressure, discomfort, or a severe intense, crushing pain which may radiate, and which may be accompanied by changes in respiration, pulse rate, and blood pressure.
The term, xe2x80x9ca condition caused by excessive calcium influx in cells in a humanxe2x80x9d includes but is not limited to conditions involving calcium influx in human cell that may be present in smooth muscle, cardiac, and other tissues including lung and brain. These conditions include, but are not limited to, cerebral ischemia, cerebral disorders such as cognitive disorders including Alzheimer""s dementia and memory impairment, arrhythmias, cardiac hypertrophy, congestive heart failure, coronary vasospasm, migraine, bronchospasm and asthma, Raynaud""s phenomenon, myocardial infarction, renal impairment and acute renal failure. The symptoms associated with these disorders include, but are not limited to, the symptoms of precordial discomfort or pain, headache, fatigue, decreased exercise tolerance, syncope, shortness of breath, nausea, lightheadedness, edema, pulmonary congestion, arrhythmia or palpitation, azotemia, and/or oliguria.
The chemical synthesis of the racemic mixture of amlodipine can be performed by the method described in Arrowsmith, J. E. et al., J. Med. Chem., 29: 1696-1702 (1986).
A technique for seperation of the amlodipine isomers from the racemic mixture is illustrated schematically (see Arrowsmith, J. E., EP 333,315) as follows:
The racemic acid 1 is converted to its cinchonidine salts in methanol solution. Upon dilution with water and standing at room temperature, a crystalline precipitate is formed which can be subsequently recrystallized to content rotation to give the diastereomerically pure cinchonidine salt 2. Further, the mother liquids from the original capitalization can be reduced in volume and stirred at room temperature, e.g. overnight, to afford a fine precipitate which can also be recrystallized to give the diastereomerically pure cinchonidine salt 2. The cinchonidine salt 2 is partitioned between ethyl acetate and dilute hydrochloric acid to liberate the acid 3. The acid 3 is then esterified using carbonyldimidazole (CDI) in near-quantitative yield by forming an imidazolide and decomposing the imidazolide with ethanolic sodium ethoxide to give 4. The azido group in 4 can then be cleanly reduced to amino by catalytic hydrogenation, giving optically pure amlodipine, which is most conveniently isolated as the salt of an acid, e.g. as the maleate 5.
The magnitude of a prophylactic or therapeutic dose of (xe2x88x92) amlodipine in the acute or chronic management of disease will vary with the severity of the condition to be treated and the route of administration. The dose, and perhaps the dose frequency, will also vary according to the age, body weight, and response of the individual patient. In general, the total daily dose ranges, for the conditions described herein, is from about 0.01 mg. to about 100.0 mg. Preferably, a daily dose range should be between about 0.5 mg to about 20.0 mg. while most preferably, a daily dose range should be between about 0.5 mg to about 10 mg. In managing the patient, the therapy should be initiated at a lower dose, perhaps about 0.025 mg to about 2.5 mg and increased up to about 20 mg or higher depending on the patient""s global response. It is further recommended that children and patients over 65 years, and those with impaired renal or hepatic function, initially receive low doses, and that they be titrated based on global response and blood level. It may be necessary to use dosages outside these ranges in some cases.
The various terms, xe2x80x9can amount sufficient to alleviate hypertension but insufficient to cause said adverse effects, xe2x80x9can amount sufficient to alleviate said condition but insufficient to cause said adverse effectsxe2x80x9d wherein said condition is angina; and xe2x80x9can amount sufficient to alleviate said condition but insufficient to cause said adverse effectsxe2x80x9d wherein said condition includes but is not limited to cerebral ischemia, cerebral disorders, arrhythmias, cardiac hypertrophy, coronary vasospasm, myocardial infarction, renal impairment and acute renal failure are encompassed by the above described dosage amounts and dose frequency schedule.
Any suitable route of administration may be employed for providing the patient with an effective dosage of (xe2x88x92) amlodipine. For example, oral, rectal, parenteral, transdermal, subcutaneous, intramuscular, and the like may be employed. Dosage forms include tablets, troches, dispersions, suspensions, solutions, capsules, patches, and the like.
The pharmaceutical compositions of the present invention comprise (xe2x88x92) amlodipine as active ingredient, or a pharmaceutically acceptable salt thereof, and may also contain a pharmaceutically acceptable carrier, and optionally, other therapeutic ingredients.
The term xe2x80x9cpharmaceutically acceptable saltsxe2x80x9d refers to salts prepared from pharmaceutically acceptable non-toxic acids including inorganic acids and organic acids.
Since the compound of the present invention is basic, salts may be prepared from pharmaceutically acceptable non-toxic acids including inorganic and organic acids. Such acids include acetic, benzene-sulfonic (besylate), benzoic, camphorsulfonic, citric, ethenesulfonic, fumaric, gluconic, glutamic, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic, mandelic, methanesulfonic, mucic, nitric, pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaric acid, p-toluenesulfonic, and the like. Particularly preferred are besylate, hydrobromic, hydrochloric, phosphoric and sulfuric acids. (See Campbell, S. F. et al., U.S. Pat. No. 4,806,557).
The compositions include compositions suitable for oral, rectal and parenteral (including subcutaneous, intramuscular, and intravenous) administration, although the most suitable route in any given case will depend on the nature and severity of the condition being treated. The most preferred route of the present invention is the oral route. The compositions may be conveniently presented in unit dosage form, and prepared by any of the methods well known in the art of pharmacy.
In the case where an oral composition is employed, a suitable dosage range for use is from about 0.01 mg. to about 100.0 mg. total daily dose, given as a once daily administration in the morning or in divided doses if required. Preferably, a dose range of between about 0.5 mg to about 20.0 mg is given as a once daily administration or in divided doses if required, and most preferably a dose range of from between about 0.5 mg to about 10.0 mg is given as a once daily administration or in divided doses if required. Patients may be upward titrated from below to within this dose range to a satisfactory control of symptoms or blood pressure as appropriate.
In practical use, (xe2x88x92) amlodipine can be combined as the active ingredient in intimate admixture with a pharmaceutical carrier according to conventional pharmaceutical compounding techniques. The carrier may take a wide variety of forms depending on the form of the preparation desired for administration, e.g., oral or parenteral (including intravenous injections or infusions). In preparing the compositions for oral dosage form any of the usual pharmaceutical media may be employed. Usual pharmaceutical media include, for example, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents, and the like in the case of oral liquid preparations (such as for example, suspensions, solutions, and elixirs); aerosols; or carriers such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents and the like, in the case of oral solid preparations (such as for example, powders, capsules, and tablets) with the oral solid preparations being preferred over the oral liquid preparations. The most preferred oral solid preparation is tablets.
Because of their ease of administration, tablets and capsules represent the most advantageous oral dosage unit form, in which case solid pharmaceutical carriers are employed. If desired, tablets may be coated by standard aqueous or nonaqueous techniques.
In addition to the common dosage forms set out above, the compounds of the present invention may also be administered by controlled release means and/or delivery devices such as those described in U.S. Pat. Nos.: 3,845,770; 3,916,899; 3,536,809; 3,598,123; and 4,008,719, the disclosures of which are hereby incorporated by reference.
Pharmaceutical compositions of the present invention suitable for oral administration may be presented as discrete units such as capsules, cachets, or tablets, or aerosols sprays, each containing a predetermined amount of the active ingredient, as a powder or granules, or as a solution or a suspension in an aqueous liquid, a non-aqueous liquid, an oil-in-water emulsion, or a water-in-oil liquid emulsion. Such compositions may be prepared by any of the methods of pharmacy, but all methods include the step of bringing into association the active ingredient with the carrier which constitutes one or more necessary ingredients. In general, the compositions are prepared by uniformly and intimately admixing the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product into the desired presentation.
For example, a tablet may be prepared by compression or molding, optionally, with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as powder or granules, optionally mixed with a binder, lubricant, inert diluent, and/or surface active or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. Desirably, each tablet contains from about 0.01 mg to about 50 mg of the active ingredient, and each cachet or capsule contains from about 0.5 mg to about 50 mg of the active ingredient, (xe2x88x92) amlodipine. Most preferably, the tablet, cachet or capsule contains either one of three dosages, 0.5 mg, 2.5 mg and 5.0 mg (as scored tablets, the preferable dose form) of the active ingredient.
The invention is further defined by reference to the following examples describing in detail the testing and preparation of the compositions of the present invention. It will be apparent to those skilled in the art, that many modifications, both to materials and methods, may be practiced without departing from the purpose and interest of this invention.

Optically pure (xe2x88x92) amlodipine compositions


There is disclosed a method of preparing a vaccine suitable for administration to humans for the prevention or treatment of cancer. The vaccine is prepared by culturing human cancer cells in a serum-free medium and recovering from the culture medium the cell surface antigens shed from the human cancer cells during culturing. After purification, the collected or recovered shed antigens are employed to produce a vaccine consisting essentially of said antigens for the treatment or prevention of human cancer.

This invention relates to human vaccines, such as vaccines for protection against pathogenic microorganisms, e.g. bacterial infections and the like, and to human anti-cancer vaccines. More particularly, and in one special embodiment, this invention relates to the preparation of human anti-cancer vaccines useful for the prevention and/or treatment of cancer, such as melanoma, breast cancer, colon cancer, lung cancer and other such cancers.
For the treatment of cancer, it has been suggested to increase tumor protective immunity by active immunization to tumor antigens, see (1) the article by R. K. Oldham entitled xe2x80x9cBiologicals and Biological Response Modifiers: Fourth Modality of Cancer Treatmentxe2x80x9d, Cancer Treat Rep (1984) :68:221-232, and (2) the article by M. J. Mastrangelo et al entitled xe2x80x9cCurrent Condition and Prognosis of Tumor Immunotherapy: A Second Opinionxe2x80x9d, Cancer Treat Rep (1984) :68:207-219. Unfortunately, this approach for the prevention and/or treatment of cancer has not been successful or completely satisfactory because of a number of problems, such as the absence in the vaccine of tumor antigens expressed by the tumor to be treated, poor characterization of the antigens in tumor vaccines, the contamination of vaccines by immunogenic but undesirable material, such as fetal calf serum (FCS) protein or transplantation antigens and additionally due to the antigenic heterogenicity of the cancer cells. Moreover, such tumor vaccines were often prepared from fresh tumor cells, the supply of which is limited so that the properties of the vaccines are not reproducible.
A clinical trial was conducted to evaluate the toxicity and immunogenicity in man of a partially purified, polyvalent, melanoma antigen vaccine and some success was indicated, see the abstract of the paper by J. C. Bystryn et al published by The Society for Investigative Dermatology, Inc. entitled xe2x80x9cPhase 1 Trial of Specific Immunotherapy of melanoma with a Polyvalent Melanoma Antigen Vaccine.xe2x80x9d
It is an object of this invention to provide an improved anti-cancer vaccine.
It is another object of this invention to provide a technique for the preparation of an improved anti-cancer vaccine.
It is another object of this invention to provide an immunotherapy for the prevention and/or treatment of human cancer.
Yet another object of this invention is to provide a technique for the production of reproducible anti-cancer vaccines.
Still another object of this invention is to provide a technique for the preparation of an anti-cancer vaccine useful when introduced into a patient to prevent and/or to treat cancer.
It is still another object of this invention to provide a technique for the preparation of vaccines for diseases caused by infectious cellular and/or subcellular organisms and/or viruses.
It is yet another object of this invention to provide a technique for the preparation of clinically or biologically important material shed from the surface of cells and the like.
How these and other objects of this invention are achieved will become apparent in the light of the accompanying disclosure. In at least one embodiment of the practices of this invention at least one of the foregoing objects will be achieved.
A human vaccine useful for the prevention and/or treatment of infections caused by pathogenic microorganisms, including viral, fungal, protozoal, amoebic and bacterial infections and the like, human cancer, including human melanoma, a skin cancer, human lung cancer, human breast cancer, human colon cancer and other human cancers is produced. For the production of a human cancer vaccine, the vaccine is produced by culturing human cancer cells, such as human melanoma cells, in a serum-free medium for the collection in the medium of cancer antigens, such as multiple melanoma associated antigens (MAAs). The vaccine produced from the shed material contains multiple cell surface antigens including tumor antigens and if prepared from cells adapted to and grown in a serum-free medium, is free of calf serum proteins. The use of bioreactors for the bulk culture of the cells and the like for vaccine production is particularly useful.
The vaccine is employed for the prevention and/or treatment of cancer in humans by administering the vaccine into the extremities of the patient a number of times a month and then once every few months thereafter for an extended period of time, such as 1-4 years, more or less. As indicated, the invention is applicable for the production of vaccines for the prevention and treatment of other cancers as well as for infectious diseases caused by bacteria, fungi, rickettsia, virus and other cellular and subcellular organisms.
The practices of this invention are hereinafter described in some detail with respect to the production of a human melanoma antigen vaccine and the treatment of melanoma patients. As indicated hereinabove, however, the practices of this invention are also applicable to the production of a human lung cancer vaccine, a human breast cancer vaccine, a human colon cancer vaccine and other human cancer vaccines as well as vaccines for infectious diseases, particularly infectious diseases caused by bacteria, fungi and other microorganisms.
In the preparation of a human melanoma antigen vaccine in accordance with this invention the vaccine was prepared from material shed by four lines of human melanoma cells: HM31, HM34, HM49, and SR-Mel-28. The cells were selected on the basis of the cells expressing different patterns of cell surface melanoma antigens and adapted to and maintained in serum-free medium, see the article by J. P. Mather et al entitled xe2x80x9cThe Growth of Mouse Melanoma Cells in Hormone Supplemented Serum-Free Mediumxe2x80x9d, Exp Cell Res 1979:120:191-200, for at least 8 weeks prior to use.
Antisera
A panel of eight murine monoclonal antibodies and two rabbit polyclonal melanoma antisera were used for MAA immunophenotyping. The antigens defined are listed in accompanying Table 1. The antigens defined by the murine and rabbit antisera are different, even though some may have similar molecular weights, as shown by variations in their distribution among melanomas (Table 3).
Vaccine Preparation
Melanoma cells were incubated at a concentration of 2xc3x97106/ml serum-free RPMI 1640 medium. After 3 hours at 37xc2x0 C., the medium was collected and the cells were removed by centrifugation at 2,000 g for 10 minutes, and larger particles were removed by recentrifugation at 12,000 g for 15 minutes. Equal volumes of medium from the four cell lines were pooled, concentrated 10-fold by vacuum ultrafiltration and made up to a final concentration. In some cases vaccines were prepared with further treatment including the addition of a non-ionic surfactant, e.g. 0.5% Nonidet P-40 (NP-40) and 0.02% sodium azide as a preservative, and then ultracentrifuged at 100,000 g for 90 minutes. The supernatant was dialyzed at 4xc2x0 C. against normal saline with 0.02% sodium azide and made up to the desired protein concentration by the addition of normal saline, passed through a 0.1 um Millex Millipore filter to remove microorganisms; and 0.5 ml aliquots dispensed into sterile, pyrogen-free glass vials. The vials were stored at xe2x88x9270xc2x0 C. until used.
A control vaccine was similarly prepared from a pool of normal peripheral leukocytes obtained from five normal individuals. Prior to use, the vaccine was tested for aerobic and anaerobic bacteria, fungi, and hepatitis antigen and also tested for mycoplasma by bisbenzamide DNA fluorochrome stain and for pyrogens by the limulus test. For MAA phenotyping, the vaccine was prepared from cells radioiodinated by the lactoperoxidase technique, see the article by J. C. Bystryn et al entitled xe2x80x9cIdentification and Solubilization of Iodinated Cell Surface Human Melanoma Associated Antigensxe2x80x9d, Int J Cancer 1977:20:165-172, and radiolabeled cells were lysed in 10 ml of 0.15 NP-40.
Assays
Protein concentration was measured by the Lowry method, see the article by O. H. Lowry et al entitled xe2x80x9cProtein Measurement with the Folinphenol Reagentxe2x80x9d, J Biol Chem 1951:193:265-275. Profiles of proteins were analyzed by SDS-PAGE and silver staining or autoradiography, see the articles, respectively, by J. C. Bystryn entitled xe2x80x9cComparison of Cell-Surface Human Melanoma Associated Antigens Identified by Rabbit and Murine Antibodies, Hybridoma 1982;4:465-472, and B. R. Oakley et al xe2x80x9cA Simplified Ultrasensitive Silver Stain for Detecting Proteins in Polyacrylamide Gelsxe2x80x9d, Anal Biochem 1980:105:361-363. Radioactivity associated with labeled macromolecules was measured by precipitation with 10% trichloroacetic acid. MAAs were assayed by protein Asepharose immunoprecipitation of labeled antigens by using the panel of antisera listed in Table 1, followed by SDS-PAGE and autoradiography. FCS proteins and Dr antigens were assayed in a similar manner by using appropriate antisera.
Clinical Studies
The vaccine was used to immunize 13 patients with metastatic malignant melanoma, see accompanying Table 4. All had intact immune function, as evidenced by skin reactivity to at least one standard recall antigen or by the ability to be sensitized to dinitrochlorobenzene (DNCB). A single lot of vaccine was used to treat all patients. Therapy was initiated at least 1 month following surgery, chemotherapy, or radiation therapy. No other therapy was given concurrently, and 0.1 ml of vaccine was administered without adjuvant into each of the four extremities weekly for eight weeks, monthly for three months, then twice at three month intervals and thereafter every six months. The initial dose of 0.25 ug/site was increased every 2 weeks to 1.0, 10.0 and 50 ug/site. Physical examination, blood count, sedimentation rate, serum chemistry profile, urinalysis, serum immunoelectrophoresis, immune complex levels, and chest x-ray were performed prior to therapy, twice monthly and every 2 months thereafter. Chest x-rays were repeated every 3 months and other radiological including as indicated. Serum for melanoma antibody was collected on two occasions prior to therapy and prior to each vaccination.
Immune Responses to Melanoma
Antibodies to melanoma surface antigens were assayed by indirect immunoprecipitation, using as antigen detergent extracts of lactoperoxidase radiodinated melanoma cells see the article by G. K. Naughton et al entitled xe2x80x9cAntibodies to Normal Human Melanocytes in Vitiligoxe2x80x9d, J Exp Med 1983:158:246-251. The cells were those used for vaccine production. Each patient served as his own control. Immunization was considered to have increased melanoma antibody level if the counts per minute (cpm) immunoprecipitated by postimmune sera were at least 50% greater than the average cpm immunoprecipitated by two preimmune sera in the same patient. The antigens defined by antibody in selected patients were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and autoradiography, see reference 11 hereinabove. Antibodies of FCS proteins in rabbit and human sera were assayed by protein A-sepharose immunoprecipitation using fetal calf serum radioiodinated with immobilized lactoperoxidase, see the article by G. S. David entitled xe2x80x9cProtein Iodination with Solid State Lactoperoxidasexe2x80x9d, Biochemistry 1974:13:1014-1021.
Cellular immunity to melanoma was evaluated by delayed-type hypersensitivity (DTH) reactions to skin tests with 10 ug of vaccine. These were considered positive if induration at least 10 mm in average diameter persisted for 24-48 hours.
Protein Profile of Vaccine
At least 20 proteins ranging in MW 30 Kd to 70 Kd were present in the vaccine as indicated by SDS-PAGE profile of proteins stained with silver in the material shed by pooled melanoma cells. Additional proteins with MWs ranging to 200 Kd could be visualized in vaccine prepared from radioiodinated cells upon SDS-PAGE profile of MAAs in vaccine prepared from radioiodinated cells immunoprecipitated by monoclonal antibodies.
To minimize the presence of FCS proteins in the vaccine, the vaccine was prepared from cells maintained in serum-free medium for at least 8 weeks. Some experiments confirmed that no detectable FCS proteins persisted in the vaccine. None could be detected by immunoprecipitation with anti-FCS serum in vaccine prepared from lactoperoxidase radioiodinated cells, even though this procedure is sensitive enough to detect FCS adhering to and shed by melanoma cells grown in the presence of serum, see the article by J. C. Bystryn et al entitled xe2x80x9cRelease of Surface Macromolecules by Human Melanoma and Normal Cellsxe2x80x9d, Cancer Res 1981:41:910-4. Two rabbits repeatedly immunized to the vaccine developed antibodies to a number of melanoma antigens in the vaccine but not to radioiodinated FCS proteins. By contrast, antibodies to FCS were readily detected in two control rabbits immunized in parallel to vaccine contaminated with trace amounts of xenogeneic serum. Finally, no antibodies to FCS were found in 13 patients repeatedly immunized to the vaccine.
To separate MAAs from transplantation antigens, shed material was treated with 0.5% NP-40 to solubilize aggregates and was subsequently ultracentrifuged. The effects of this procedure on proteins, tumor, and transplantation antigens are summarized in Table 2. Ultra-centrifugation of shed material not treated with detergent removed much of the radioactivity associated with labeled macromolecules, in addition to those MAAs defined by polyclonal antiserum SB29 and 100% of those associated with Dr antigens. Ultracentrifugation of detergent-treated shed material still resulted in a loss of all detectable Dr antigens but reduced the loss of labeled macromolecules to 17% and of MAAs to 30%. Thus, detergent treatment and ultracentrifugation removed Dr antigens with an acceptable loss in MAAs.
Antigenic Properties of Vaccine
Vaccine prepared from radioiodinated cells was immunophenotyped with a panel of 10 melanoma antisera. The results are summarized in Table 1. Most of the MAAs tested were present in the vaccine. Three batches of vaccine prepared several months apart all contained the MAAs tested, see accompanying Table 1. Upon freeze-thawing three times or storage at xe2x88x9270xc2x0 C. for four months, there was no loss in antigenic activity.
Distribution of MAAs in Various Melanomas
Because it is desirable that the vaccine contain at least one tumor antigen which will be present on most of the melanoma tumors to be treated, it was examined whether or not the panel of MAAs in the vaccine satisfied this requirement. Fifteen melanomas were lactoperoxidase radioiodinated and immunophenotyped for the MAAs present in the vaccine. There were marked differences, see accompanying Table 3, in the pattern of MAAs expressed by each melanoma. However, all the melanomas expressed several of the MAAs present in the vaccine.
Phase I Trial
Thirteen patients with metastatic melanoma were immunized with increasing doses of vaccine for 2 months, see accompanying Table 4, and 10 of these received 2 to 10 additional booster doses during the period 5 months to 2 years. No toxicity was observed other than transient urticaria at the site of injection. A similar urticarial reaction occurred in the skin of each of five patients tested with sterile saline containing an amount of NP-40 equal to that present in the vaccine. There was no reaction to skin tests to sterile saline alone.
Additional Clinical Trials
Clinical studies were performed to determine the toxicity of the vaccine in patients with early melanoma and the effect of several immunization strategies on immunogenicity. A total of 55 melanoma patients (36 with Stage II disease metastatic to regional nodes and 19 patients with Stage III widely disseminated disease) have been treated. There was no toxicity other than transient inflammation at the site of vaccine injection.
Immune Response to Immunization
Antibodies to surface antigens on melanoma cells, assayed by indirect immunoprecipitation, increased in five (38%) patients, see accompanying Table 5. The increase was usually small, although in one patient (patient 1) peak binding activity was 300% greater than the preimmune level. Antibody levels increased in some patients within 1 week following the first immunization, suggesting that a secondary type of immune response was elicited. The immunogenic antigens that stimulated the production of melanoma antibodies were identified in three patients by SDS-PAGE and autoradiography. These were proteins with MWs of about 75, 85, and 200+ Kd. The larger antigen appeared to be more immunogenic, since all three patients developed antibodies to it, whereas antibodies to the other two antigens were present in only one patient. None of these antigens was related to FCS proteins, since their binding was not blocked by excess cold FCS. Furthermore, after 2 months of immunization, there was no increase in antibodies to FCS in any of the patients, see accompanying Table 6.
Cellular immunity to melanoma, evaluated by DTH reaction to skin tests with the vaccine, was induced in four (31%) patients, see accompanying Table 5. Skin tests were negative prior to immunization in all patients. Three to six immunizations with 10 ug or more of vaccine were required to induce a positive reaction. There was no reaction in positive patients to skin tests with equal amounts of human albumin or to a control vaccine prepared from pooled normal peripheral leukocytes. Overall, there was an increase in humoral and/or cellular immune response to melanoma following immunization in eight (62%) of the patients.
In subsequent clinical trials it was found that antibody and/or cellular immune responses to melanoma were induced more frequently in Stage II (69% of 36 patients) than in Stage III (53% of 19 patients) disease. The ability of different immunization schedules, alum or pretreatment with low dose cyclophosphamide, to potentiate immunogenicity was compared after 2 months of immunization. Bi-weekly immunization with a fixed intermediate dose of vaccine was more immunogenic than weekly immunization with escalating vaccine doses. Alum increased slightly the intensity of cellular responses while pretreatment with cyclophosphamide augmented slightly both the incidence and intensity of cellular immune responses. There was a reciprocal relationship between the induction of humoral and cellular immune responses. The most effective immunization schedule consisted of pretreatment with cyclophosphamide which augmented antibody and/or cellular immune responses to melanoma in 83% of patients.
The vaccine is capable of augmenting immune responses to a patient""s own tumor. There were examined the density and distribution of lymphocytic inflammatory cells in cutaneous metastases that developed during the course of vaccine immunization in 11 patients and comparison was made in a blinded fashion to the lymphocytic infiltrate present in similar cutaneous melanoma metastases removed from 22 non-immunized, randomly selected patients. Dense cellular infiltrates ( greater than 15 lymphoid cells/high power field HPF) were more frequent in tumor nodules of immunized patients (10 of 11 nodules, 91%) than in those of control patients (9 of 22 nodules, 41%), p 0.01. The cells were more likely to be infiltrating within the tumor nodules in immunized patients (7 of 11 nodules, 64%) than in control patients (5 of 22 nodules, 23%), p 0.05. The incidence of lymphoid cells at the periphery of nodules was similar in both groups.
These results indicate that immunization to melanoma vaccine can augment lymphocyte infiltration into tumors in vivo, and suggests that polyvalent allogeneic melanoma vaccines can boost a patient""s ability to mount an immune response to his own tumor.
Effect of Immunization on Tumor Growth
In the initial Phase I study, two of the 13 immunized patients have done well, see accompanying Table 4. There was a complete remission in a 46-year-old male (patient 10) who developed three disseminated cutaneous metastasis one year following excision of a melanoma of the right upper back. Two metastases were removed and the third left as a marker. It increased in size 30% during the first months of immunotherapy, was stable for several months, and then slowly decreased in size by 50%. It was removed 10 months after onset of immunotherapy and histologically was a regressed malignant melanoma with only pigment-laden macrophages at the site. The patient is currently disease-free, 36 months after onset of immunotherapy.
There was a long-term stable disease in a 75-year-old woman (patient 9) who developed two cutaneous metastases 2 years after excision of a malignant melanoma of the right calf. These were removed, and she was treated with dimethyltriazenoimidozole carboxamide (DTIC) and actinomycin D. Three years later, six new cutaneous metastases appeared, and she was started on vaccine immunotherapy. Cutaneous metastases increased in number and size during the next 4 months until she had 15. She was continued on immunotherapy, and there has been no further increase in size or number of metastases. The largest lesion regressed spontaneously 6 months after onset of immunotherapy. The patient currently has stable disease 14 months after onset of immunotherapy.
In other studies involving 55 patients with stage II melanoma, it was found that recurrence of melanoma was less common in patients with a cellular immune response to the vaccine (35%) than in those with an antibody response (86%) p less than 0.05 and the disease free interval was prolonged from about 39 to greater than 100 weeks. These results suggest that active immunization to the melanoma vaccine can slow the progression of melanoma in some patients.
Several features of the technique used to prepare the vaccine deserve attention, because they were designed to minimize problems. The vaccine was prepared from melanoma cells grown in serum-free medium to avoid contamination with FCS proteins. These proteins are highly immunogenic and responsible for most immune responses induced by human tumor vaccines prepared from cultured cells, see the articles by P. O. Livingston et al entitled xe2x80x9cSerological Responses of Melanoma Patients to Vaccines Derived from Allogenic Cultured Melanoma Cellsxe2x80x9d, Int J Cancer 1983:31:567-575, P. O. Livington et al entitled xe2x80x9cSerological Response of Melanoma Patients to Vaccines Prepared from VSV Lysates of Autologous and Allogenic Melanoma Cellsxe2x80x9d, Cancer 1985:55-713-720 and M. S. Mahaley, Jr. et al entitled xe2x80x9cImmunobiology of Primary Intracranial Tumors. Part 8: Serological Responses to Active Immunization of Patients with Anaplastic Gliomasxe2x80x9d, J Neurosurg 1983,59:208-216. This approach appears to have been successful, since no FCS proteins were detectable in the vaccine by direct assay or by the more sensitive method of looking for an immune response to these proteins in animals or persons immunized to it.
Cultured cells were used to ensure a continued and reproducible source of material, a basic requirement for vaccine development, see reference article 1. Three batches of vaccine prepared several months apart all contained the MAAs that were tested for, indicating that vaccine containing similar antigens can be made reproducilibly. Quantitative changes in the amount of each antigen in the vaccine have not been excluded.
The vaccine was prepared from a pool of cells, selected because they expressed different patterns of surface MAAs. This was done to create a polyvalent vaccine, which is desirable in treating melanoma cells (and, indeed, for treating other cancer cells) that are antigenically heterogeneous, see the articles by J. C. Bystryn et al entitled xe2x80x9cImmunophenotype of Human Melanoma Cells in Different Metastasesxe2x80x9d, Cancer Res, in press, 1985, M. Y. Yeh et al entitled xe2x80x9cClonal Variation in Expression of a Human Melanoma Antigen Defined by a Monoclonal Antibody, J Immunol 1981:126:1312-7, A. P. Albino et al entitled xe2x80x9cHeterogeneity in Surface Antigen and Glycoprotein Expression of Cell Lines Derived from Different Melanoma Metastases of the Same Patientxe2x80x9d, J Exp Med 1981:154-1764-1778. The vaccine contained multiple MAAs. Most of the MAAs tested for were present in the vaccine; it is therefore likely that the vaccine contains additional MAAs that were not tested for. At least one of the MAAs present in the vaccine was expressed by each of 15 metastatic melanomas that were immunophenotyped. Thus, a mixture of tumor antigens in the vaccine would be appropriate to circumvent antigenic heretogeneity among melanomas.
Shed antigens were used because they are partially purified. They are separated from the bulk of cytoplasmic components, which are slowly released ( less than 5%) during the short collection period; surface components, however, are released rapidly, see the article by J. C. Bystryn et al entitled xe2x80x9cShedding and Degradation of Cell-Surface Macromolecules and Tumor-Associated Antigens by Human Melanomaxe2x80x9d. In: Reisfeld R. A., Ferrone S., eds. Melanoma Antigens and Antibodies, New York: Plenum Press. 1982:37-52. In addition, shed material is enriched in surface antigens which are more likely to be relevant for immunotherapy and can be repeatedly harvested from the same cells, thus reducing culture requirements.
Finally, detergent treatment and ultracentrifugation were used to separate tumor from Dr antigens. Much of the material shed by melanoma cells is contained in fragments that are sedimented by ultracentrifugation. Treatment with detergent breaks up these fragments so that most tumor antigens remain in suspension following ultracentrifugation, however, as found and confirmed by others, see the article by R. A. Reisfeld et al entitled xe2x80x9cApproaches for the Isolation of Biologically Functional Tumor-Associated Antigensxe2x80x9d, Cancer Res 1977:37:286-2865, ultracentrifugation still causes sedimentation of detergent-treated transplantation antigens. This consequently provides a simple way of separating transplantation from tumor antigens.
The vaccine produced was found to be safe and immunogenic in 55 patients with metastatic melanomas. There was no toxicity other than transient urticaria at the site of the injection. Immune responses to melanoma were induced or augmented in 64% of the patients. Cellular immunity was induced in 51% of the patients. me reaction seemed directed to melanoma, since there was no response to concurrent skin tests with equal amounts of control vaccine prepared from pooled allogenic peripheral leukocytes or to human albumin. The cellular response was induced, rather than preexisting or due to an irritant in the vaccine, since there was no reaction to the initial injections of vaccine at doses that subsequently led to strong skin reactions.
Humoral immunity to melanoma was increased in 24% of patients. me responses were generally weak; however, a major immunogenic contaminant in prior vaccine prepared from cultured cells, i.e. FCS proteins, see the article by H. F. Seigler et al entitled xe2x80x9cSpecific Active Immunotherapy for Melanomaxe2x80x9d, Ann Surg: 1979:190:366-37, appears to have been successfully removed. No patient developed FCS antibodies, and by blocking studies with cold FCS, the melanoma antibodies induced in some patients were not directed to FCS. The immunogenic antigens in the vaccine were surface proteins with MWs of about 75, 85, and 200+ Kd. As determined by migration on SDS-PAGE, these were different from HLA or Dr antigens, whose MW is 50 Kd.
Three of the 19 patients with Stage III disease did well following immunization. One had complete biopsy proven regression of metastatic cutaneous disease and remained healthy and free of melanoma 35 months after starting immunization. Another remained disease-free following resection of a solitary gall bladder metastases 13 months after starting vaccine therapy. The third patient showed a dramatic slowing of previously rapidly progressing cutaneous disease following initiation of vaccine immunotherapy of progression of cutaneous metastases resumed after 6 months but the patient remained otherwise healthy and fully active 25 months after starting immunization.
These results are unexpected since immunotherapy is not thought to be effective in patients with advanced diseases, see the article by R. K. Oldham et al entitled xe2x80x9cImmunotherapy of the Old and the Newxe2x80x9d, J Biol Response Mod 1983.2:295-309. However, because the number of patients is small and the course of melanoma is usually unpredictable, it is difficult to judge the significance of these favorable responses in patients with advanced melanoma.
In patients with less advanced Stage II melanoma, the vaccine appears to be capable of slowing the progression of melanoma in some cases. Recurrences were lower and disease free interval was prolonged in 20 of 45 patients with Stage II melanoma who developed a cellular immune response to the vaccine. These differences were statistically significant.
The availability of a characterized polyvalent melanoma antigen vaccine that can be reproducibly made, safe to use, and immunogenic, should permit a systematic study of the factors that influence the effectiveness of active, specific immunotherapy of melanoma.
Other details of the practices of this invention described hereinabove are to be found in the article by J. C. Bystryn et al entitled xe2x80x9cPreparation and Characteristics of a Polyvalent Human Melanoma Antigen Vaccinexe2x80x9d, Journal of Biological Response Modifiers 1986:5:211-224. Also of interest for more details of the practices of this invention is the to be published article by J-C Bystryn et al entitled xe2x80x9cImmunogenecity of a Polyvalent Melanoma Antigen Vaccine in Manxe2x80x9d, the article by M. Dugan et al entitled xe2x80x9cRelationship between Immune Responses to Melanoma Vaccine Immunization and Tumor Regression in Manxe2x80x9d, Kaplan Cancer Center, NYU School of Medicine, New York, N.Y., published in American Federation for Clinical Research, 1987 and the article by R. Orate et al entitled xe2x80x9cInduction of Lymphocytic Cell Infiltrate on Human Melanoma Nodules by Active Immunization to Melanoma Antigen Vaccine n to be published by AACS 1987.
In the description of the invention set forth hereinabove, emphasis has been placed upon the preparation of a vaccine useful for the prevention and/or treatment of melanoma. It is clearly indicated, however, that the concept and practices of this invention are generally applicable to the preparation of vaccines to treat or prevent human cancers and for the preparation of vaccines to treat or prevent infectious diseases in man and animals.
The subject invention involves in a special embodiment, an immunotherapeutic approach for the prevention and/or treatment of cancer and infectious diseases. The administration to a patient of a vaccine in accordance with this invention for the prevention and/or treatment of cancer can take place before or after a surgical procedure to remove the cancer, before or after a chemotherapeutic procedure for the treatment of cancer, and before or after radiation therapy for the treatment of cancer and any combination thereof. It would appear, therefore, that the cancer immunotherapy in accordance with this invention would be a preferred treatment for the prevention and/or for the treatment of cancer, particularly since the risk and side effects involved are substantially minimal compared with the other treatments, e.g. surgery, chemotherapy and radiation therapy. A unique aspect of this invention is that the vaccines have the potential or capability to prevent cancer in individual s without cancer but who are at risk of developing cancer.
The administration of a cancer vaccine prepared in accordance with this invention, is generally applicable to the prevention or treatment of cancer. Cancers which could be suitably treated in accordance with the practices of this invention include cancers of the lung, breast, ovary, cervix, colon, head and neck, pancreas, prostate, stomach, bladder, kidney, bone liver, esophagus, brain, testicle, uterus and the various leukemias and lymphomas.
The vaccines in accordance with this invention, like the above-described melanoma vaccines, would be derived from the tumor or cancer cells to be treated. For example, in the treatment of lung cancer in accordance with the practices of this invention, the lung cancer cells would be treated as described hereinabove to produce a lung cancer vaccine. Similarly, breast tumor or cancer vaccine, colon cancer vaccine, pancreas cancer vaccine, stomach cancer vaccine, bladder cancer vaccine, kidney cancer vaccine and the like would be produced and employed as immunotherapeutic agents in accordance with the practices for the prevention and/or treatment of the tumor or cancer cell from which the vaccine was produced.
Vaccines in accordance with this invention could, as stated, also be prepared to treat various infectious diseases which affect man and animals by collecting the relevant antigens shed into the culture medium by the pathogen. As there is heterogenecity in the type of immunogenic and protective antigens expressed by different varieties of organisms causing the same disease, polyvalent vaccines could be prepared by preparing the vaccine from a pool of organisms expressing the different antigens of importance.
In the practices of this invention set forth hereinabove for the treatment of melanoma, the vaccine was intradermally or subcutaneously administered to the extremities, arms and legs, of the patients being treated. Although this approach is generally satisfactory for melanoma and other cancers, including the prevention or treatment of infectious diseases, other routes of administration, such as intramuscularly or into the blood stream may also be used. In addition, the vaccine can be given together with adjutants and/or immuno-modulators to boost the activity of the vaccine and the patient""s response.
The vaccines of this invention, as described hereinabove, have been prepared from cells grown in flasks. The production of the vaccine can, if desired, be scaled up by culturing the cells in bioreactors or fermentors or other such vessels or devices suitable for the growing of cells in bulk. In such apparatus the culture medium would be collected regularly, frequently or continuously to recover therefrom the cell-shed or tumor-shed or materials or antigens before such materials or antigens are degraded in the culture medium.
If desired, devices or compositions containing the vaccine or antigens produced and recovered in accordance with this invention and suitable for sustained or intermittent release could be, in effect, implanted in the body or topically applied thereto for the relatively slow release of such materials into the body.
In summary, the essential features of vaccine preparation in accordance with this invention are the following:
(a) The use of cultured cells to provide a continued and reproducible supply of material for vaccine production.
(b) The use of a pool of cell lines. Each line is selected on the basis of it expressing a different pattern of cell surface antigens. This is done to ensure that the vaccine contains a broad representation of different antigens, hence, circumventing the problem antigenic heterogeneity of tumor cells.
(c) Adapting and maintaining the cells in serum-free medium. This excludes from the vaccine undesirable and highly immunogenetic fetal cal f serum proteins shown to be free of hepatitis, AIDS, and other undesirable pathogens.
(d) Using shed material to obtain antigens for vaccine preparation. This provides numerous advantages. It greatly simplifies the process of collecting bulk, on-line, production of antigens. The antigens which are collected are solubilized. They are also partially purified as they are separated from the bulk of cytoplasmic cellular components which are shed much more slowly. This greatly simplifies subsequent purification or other biochemical procedures. Lastly, shed material permits the antigens to be harvested from the cells without cell destruction. This greatly reduces the cost of cell culture requirements and hence the time and cost of making the vaccine. The continued use of the same cells to generate antigens also greatly reduces cell culture requirements.
(e) The culturing operation for the preparation of the vaccine, particularly the production of the cancer associated cell surface antigens wherein the cancer cells are cultured is carried out for a suitable period of time, such as for a period of time in the range 2-6 hours or higher, such as in the range 12-24 hours, more or less. Desirably, the culturing operation is carried out for a period of time before the shed antigens are substantially degraded in the culture medium and especially whereby the shed antigens are partially separated from the bulk of the cytoplasmic cancer cell cellular components which are shed more slowly. As would be apparent to those skilled in the art, there are degrees of flexibility with respect to the culturing operation for the production of the shed antigens.
The above steps in vaccine preparation can be individualized to satisfy the requirements of particular vaccines. In the case of melanoma vaccines the shed material is concentrated and in some cases treated with detergent and ultracentrifuged to remove transplantation alloantigens.
The disclosures of all the above-identified reference publications are herein incorporated and made part of this disclosure.


Embodiments of the present invention provide users with suggested content that is weighted based on the social network context of the suggestion. In particular, the suggested content is selected based on incorporating the preferences of users having a relationship with the user. For example, content recommendations from a family member or known friend of the user may be highly weighted over other recommendations.

In its short history, Internet usage has been mainly driven by portals and search engines, such as Yahoo! and Google. Recently, the rapid growth of social networking sites, such as MySpace and Facebook, has revealed a new trend of Internet usage. Social networking generally relates to services and tools that help users maintain and expand their circles of friends usually by exploiting existing relationships. Social networking sites have shown potential to become the places on the Internet where many people spend most of their time, thus making these sites the main entry point for online activity. Often times, these social networking sites can become the focal point of sharing information, such as links, multimedia, music, and the like.
In general, social networking sites and other online services of the Internet offer a mix of features and tools, such as message boards, games, journals or web logs ("blogs"). Many of these sites try to build communities around multi-media or popular culture, such as television, film, music, etc. These sites and their features are designed to keep users clicking on advertising-supported pages of the site. Thus, the known social networking sites employ a closed platform of services that attempt to keep their user-base captive to the site.
One feature of social networking sites is providing recommendations for content, such as video, audio, images, etc. Unfortunately, given the proliferation of social networking sites and their membership, it can be difficult to trust their recommendations. In addition, often times the recommendations are of little or no interest to the user.
Accordingly, it would be desirable provide methods and systems for providing relevant recommendations for content.

Automatic generation of content recommendations weighted by social network context


The present invention 10 discloses a stopwatch 19 seated in a molded cavity 36 of a compressible foam-like block of material 18 enclosed by a cover 20 having a transparent element 22 to hold the stopwatch 19 within the cavity wherein the block of material is mounted to the handlebars 16 of a motorcycle. The stopwatch 19 has electrical connection means 24 with an on/off switch 26 that is mounted near the handlebar grip 28 whereby the stopwatch can be started and stopped by the rider using their thumb. Positioned within the top of the material 18 is the cavity 36 having an angular base for the placement therein of a timepiece 19 using mating hook and loop material 32 wherein the angular base positions the timepiece at an angle so that the timepiece is easily viewable by the rider. Also shown is a protective cover element 20 having mating hook and loop material 30 thereon forming means for closure that is wrapped around the foam-like material 18. The timepiece 19 may also have a lighted face for use in the dark.

1. Field of the Invention
The present invention relates generally to time pieces and, more specifically, to a stopwatch enclosed within a housing that is mounted to the handlebars of a motorcycle having a switch mounted on either side of handlebars for starting and stopping the stopwatch.
The handlebar stopwatch provides means for a motorcycle rider to accurately time himself or herself during practice by pressing a switch mounted to handlebars for starting and stopping the stopwatch. It can also be used when riding for exercise by setting it on 15, 30, 45 minute extension intervals. Riders can also compare times with each other and see where they stand in comparison with the track record or their best time. Riders should be able to scroll through each lap time by pressing another button on the time piece.
2. Description of the Prior Art
There are other timing devices designed for time keeping. Typical of these is U.S. Pat. No. 490,750 issued to Kuhn on Jan. 31, 1893.
Another patent was issued to McGriff on Aug. 8, 1893 as U.S. Pat. No. 503,058. Yet another U.S. Pat. No. 1,262,040 was issued to Hall on Apr. 9, 1918 and still yet another was issued on Feb. 27, 1923 to Bush as U.S. Pat. No. 1,446,613.
Another patent was issued to Davies on Mar. 24, 1964 as U.S. Pat. No. 3,125,884. Yet another U.S. Pat. No. 3,708,750 was issued to Bucks, et al., on Jan. 2, 1973. Another was issued to Wilson, et al., on Dec. 25, 1979 as U.S. Pat. No. 4,179,877 and still yet another was issued on Jun. 24, 1986 to Adkins as U.S. Pat. No. 4,596,370.
Another patent was issued to Mire on Oct. 21, 1986 as U.S. Pat. No. 4,618,081. Yet another U.S. Pat. No. 4,819,217 was issued to Houlihan on Apr. 4, 1989. Another was issued to Rogowski on Jan. 1, 1991 as U.S. Pat. No. 4,981,243 and still yet another was issued on Apr. 30, 2002 to Lee as U.S. Pat. No. 6,378,815.
The combination with the handle rod C, of the two-part clamp detachably secured thereto, the flat spring arm E having a portion lying flat on and held to the tipper part of the said clamp and curved as shown and its other end curved and extended at right angles to the length of the arm to conform to the curvature of a clock and to partially embrace the same, and the clock sander which the spring arm passes and to which it is secured, substantially as shown and described.
An attachment to a bicycle for holding articles, consisting of a device constructed of wire or other suitable material and having a frame to embrace the handle of the bicycle and a clip to embrace the post or standard thereof, said clip terminating in screw threaded extensions, and a clamping plate and thumb-nuts for forcing the plate against the post or standard, substantially as and for the purpose set forth.
Means for indicating the elapsed time during which a machine is in motion, comprising a clock mechanism, a shaft driven by the machine, a friction clutch having one member mounted on said shaft, a detent device for said clock mechanism operatively connected with the other clutch member; and a coiled spring normally holding said decent device in operative position.
In a clock stopping device to cooperate with a vehicle, the combination of a member adapted to, be actuated through the agency of the running gear of the vehicle, yielding stop mechanism for stopping the clock, and means positively actuated by said member and engaging the stop means to actuate to actuate the same.
An elapsed-time testing apparatus for association with a vehicle, said apparatus comprising a road engaging wheel other than a wheel of the vehicle and provided with means far ready attachment to and detachment from the vehicle, traversal responsive means comprising a screw shaft and means for communicating motion from said wheel to said shaft, traveling nut means operably associated with said shaft and traversable therealong between predetermined limits, a normally closed switch cooperating with said traveling nut means, a normally open second switch carried on said apparatus, said second switch balanced to readily respond to the inertial force exerted by said vehicle from a standing start, an electrically controlled time indicating instrument, and an electrical circuit including said instrument and said switches in series, whereby in response to the initial forward lurch of said vehicle from a standing start said second switch will be operated to its circuit closing position to cause the operation of said time indicating instrument until said first recited switch is operated to its open circuit position by said traveling nut means to terminate operation of said time indicating instrument to indicate the elapsed time of traversal of said vehicle over a measured course
An electronic apparatus for measuring the time required for a vehicle to travel a predetermined distance such as a quarter mile. The exemplary embodiment described comprises pulse generating means for producing a pulse for each increment of distance traveled in combination with an integrating means for accumulating such pulses and producing a stop count signal when the output of the integrating means reaches a predetermined level. By adjusting a duty cycle of the individual pulses input to the integrating means, the predetermined point at which the stop count output is produced may be made to correspond to a desired predetermined value of traveled distance. Clock pulses are counted by a digital counter from a time determined by either manual or automatic start means until said stop count signal is produced at which time the counter is adapted to hold its contents until being reset. An indicator is also associated with the counter for visually displaying the contents of the counter indicating the elapsed time required for the vehicle to travel the predetermined distance as determined by the pulse width adjustment.
A mounting structure incorporated into the dashboard of an automotive vehicle or the like for removably supporting a portable, self-contained, battery operated alarm clock to enable the clock to be easily mounted in the dashboard and just as easily removed therefrom to enable the clock to be used in the vehicle in a customary manner and removed therefrom and carried to any desired location for use as a portable alarm clock having general utility. The clock is especially useful by travelers since it can be taken out of the automobile and into the motel or hotel room so that it can be used as an alarm clock. The mounting structure and clock are uniquely associated so that the clock may be easily inserted but yet securely retained in the vehicle dashboard even when traversing rough or uneven roadways and the like with a manually manipulated structure enabling easy removal of the clock from its secure position in the dashboard.
A container holder for clamping to a tubular support member. The container holder comprises a base member for supporting on its upper surface a tubular container open at its upper end. The lower surface of the base member includes an adjustable clamp for adjustably gripping different sized tubular supports. The container holder is either formed of rigid or flexible insulating material. When rigid, the container holder, base and adjustable clamp are formed as an integral unit. When formed of flexible insulating material, the container holder can be fixedly attached to the upper surface of the base member or removably attached by means of Velcro.RTM. hook-and-eye fastening material. The clamping means may be located on the rigid container intermediate its ends.
Apparatuses for mounting flashlights to bicycles have concave surfaces for directly supporting flashlights and which are releasably held thereto by Velcro type material bearing straps. The apparatuses also have semi-cylindrical channels adapted to the placed in mating contact with bar elements of the bicycles and releasably held thereto by other Velcro type material bearing straps.
A bicycle handlebar holder for a wrist instrument with pushbuttons and electrical contact means, said holder having a holder body, a cover which is pivotable with respect to the body and defining therewith a receptacle for holding the case of said wrist instrument when the cover is closed, a clamp for attaching the body to a bicycle handlebar, a plurality of electrical contacts disposed in the body and opening into the receptacle, and at least one manual actuator disposed in the cover and adapted to be operated from outside of the cover and having a portion movable inside said receptacle which is disposed to depress a pushbutton when the actuator is operated.
An audio equipment carrier assembly to be mounted on the handlebars or other like support members of various two and three wheel vehicles. The preferred embodiment includes a carrier assembly body adapted to receive and contain a Walkman-style personal radio having headphones which are very popular. Bicyclists and the like which desire to listen to music while riding may use the equipment carrier of the present invention to hold the radio or other audio equipment on the front of the bicycle while wearing the headphones for the equipment, which is connected to the audio source.
The mounting assembly includes a shaft having a planar member mounted on a top end of the shaft. The planar member is preferably mounted so it pivots with respect to the shaft. The shaft is insertable in a hollow cylinder on a motor vehicle. In the case of a motorcycle, a portion of the shaft is inserted in a steering stem of a steering head. A plurality of o-rings, attached to a portion of the shaft inserted in the cylinder, secure the mounting assembly in place. A Velcro TM. piece, fixed to a top surface of the planar member, clasps to a corresponding Velcro TM. piece of an electronic device placed on the mounting assembly.
While these timing devices may be suitable for the purposes for which they were designed, they would not be as suitable for the purposes of the present invention, as hereinafter described.


A dock leveler that includes a pivotal ramp with a moveable lip for loading and unloading a truck at a loading dock also includes a lip seal that helps seal an air gap along the side edges of the lip. When the back end of the truck is up against a dock bumper, the lip helps block off an air gap that spans an area generally defined by the rear edge of the truck, an inside edge of the bumper, and the side edge of the lip that generally faces the bumper. When the lip moves from its extended, operative position to its stored position, the lip forces the lip seal to move underneath the deck. The lip seal is biased to an operative, sealing position by a spring or a counterweight, so as the lip extends to its operative position, so does the lip seal.

1. Field of the Invention
The subject invention generally pertains to loading docks, and more specifically to a seal that seals against a dock leveler lip.
2. Description of Related Art
A typical loading dock of a building includes an exterior doorway with an elevated platform for loading and unloading vehicles, such as trucks and trailers. To compensate for height differences between the loading dock platform and an adjacent bed of a truck or trailer, many loading docks have a dock leveler. A typical dock leveler includes a deck, also known as a ramp or dockboard, which is pivotally hinged along its back edge to vary the height of its front edge. An extension plate, or lip, extends outward from the deck""s front edge to span the gap between the rear of the truck bed and the front edge of the deck. Depending on the particular dock leveler, some lips move linearly and others pivot between a stored position and an extended, operative position. In moving to the stored position, the lip usually retracts to where it does not interfere with a vehicle backing into the dock. In the extended, operative position, the lip extends from the deck""s front edge and rests upon the truck bed to form a bridge between the two. This allows personnel and material handling equipment to readily move on and off the vehicle during loading and unloading operations.
To protect the building and the dock leveler from direct vehicle impact, loading docks often include bumpers. Bumpers also help establish a predetermined distance between the rear of the vehicle and the dock leveler, so the dock leveler can first raise and then lower its lip upon the rear of the vehicle with a safe amount of overlap, or lip purchase, between the lip and the bed of the vehicle. Bumpers are usually installed near the bottom of the doorway, adjacent either side of the dock leveler lip and protrude a few inches out from the face of the dock, where they can be abutted by the rear of the vehicle.
To help block out rain, snow and outside air from entering the building through air gaps between the back of the vehicle and the exterior face of the building, usually either a dock seal or a dock shelter is installed around the perimeter of the doorway.
Dock shelters often have projecting members that protrude outwardly from the face of the building and run generally along the top and lateral sides of the doorway. From a protruding edge of the projecting members, a top extending member and two laterally extending members may extend inward and generally parallel to the building face to help seal against the truck""s top and sides, respectively. The laterally extending members are often made of a fabric or flexible foam. Two inner bottom corners of the dock shelter at the lower back sides of the truck are often partially sealed by draft pads attached to the lower ends of the projecting members. The dock leveler lip resting upon the rear of the vehicle is often relied upon to seal most of the doorway""s lower edge.
Typical dock seals comprise a resiliently compressible foam core protected by a fabric outer cover. They are usually mounted to the face of a building, along the top and both sides of the doorway. With dock seals, sealing is provided by backing the truck directly up against the seal. The seal then compressively conforms to the shape of the truck""s rear edges. The foam core provides the necessary compliance and resilience to repeatedly conform to the shape of various trucks, while the outer cover protects the foam core from dirt, water and abrasion. As with dock shelters, dock seals also rely on the dock leveler lip to seal most of the doorway""s lower edge.
With conventional dock seals and dock shelters, side clearance between the lip of the dock leveler and the inner side of each bumper creates areas that are difficult to seal off. Air gaps in these areas can cause a terrible upward draft into the building, particularly if the interior of the building is at a lower pressure than the outside air. The air gaps can be especially large for a dock leveler with a tapered lip whose width across the doorway is narrower along the tip or distal edge of the lip than along a proximal edge where the lip is hinged to the deck. Tapered lips create gaps that are similarly tapered (e.g., triangular or trapezoidal). Regardless of the air gap""s shape, sealing such openings can be difficult to do, because the lip must be able to move relative to the bumper. The lip not only needs to move between a stored and an extended position, but also needs to move vertically with the up and down pivotal movement of the dock leveler""s deck. Due to the difficulty of sealing the air gaps between a dock leveler lip and adjacent bumpers, such air gaps are known to be left open.
In some embodiments, a lip seal extends between a bumper of a loading dock and a lip of a dock leveler.
In some embodiments, a lip seal engages a bumper of a loading dock and a lip of a dock leveler.
In some embodiments, a lip seal engages the underside surface of a dock lever lip.
In some embodiments, a moveable seal member engages the bumper of a loading dock.
In some embodiments, a seal member is pivotal about an axis disposed underneath the deck of a loading dock.
In some embodiments, a seal member is adapted to engage a vehicle, the lip of a dock leveler, and the bumper of a loading dock.
In some embodiments, a seal member is able to seal a gap existing along a tapered edge of a dock leveler lip.
In some embodiments, a seal member for a dock leveler lip includes a compressible foam core with a pliable cover.
In some embodiments, a seal member includes a plurality of tabs adapted to engage an inside surface of a bumper.
In some embodiments, a seal member is adapted to engage the lip of a dock leveler over an area of contact that moves as the lip moves relative to the dock leveler""s deck
In some embodiments, a seal member is adapted to engage the bumper over an area of contact that moves as the pivotal deck pivots
In some embodiments, a spring urges the seal member to an operative position.
In some embodiments, a counterweight urges the seal member to an operative position.