JP2008540428A - Conjugated aromatic compounds for diagnosis and treatment - Google Patents

Abstract

Disclosed herein are compounds, compositions, and methods of using such compounds and compositions for the treatment of hyperproliferative conditions, including precancerous and cancerous conditions. In some embodiments, the compound comprises a resonance modifying compound that can be utilized in this manner as a coupling agent that can interact with the immune system to monitor or promote immune function. Such resonance modifiers have unique electromagnetic properties that attract immune cells to the target site to which the resonance modifier is applied. The electromagnetic properties (such as voltage amplitude) of the target site are altered by the presence of the resonance modifier and are indicative of immune function. The addition of an external stimulus (such as an applied electromagnetic field) can also enhance the immune stimulating and triggering properties of the resonance modifier. Specific examples of resonance modifiers are disclosed, and these materials generally have a resonant intramolecular dipole moment that is capable of electrostatic interaction with the biological environment. In certain embodiments, the compound aggregates at the treatment site to enhance therapeutic activity. In other examples, the compound promotes platelet production.

Description

[Description of government support]
The US government has certain rights in this invention. Several studies on the development of the present invention were conducted under National Cancer Institute grant number R43 CA89772-01.
[Cross-reference to related applications]
This application claims priority from US Provisional Application No. 60 / 678,089 filed May 4, 2005, which is hereby incorporated by reference in its entirety.
[Technical field]
The present invention relates to a method for monitoring and correcting immune function, for example in the diagnosis and / or treatment of infectious and neoplastic diseases. The present invention further discloses methods of treating lesions, such as hyperproliferative lesions associated with infectious pathogens and / or inflammation.

  Immunotherapy involves modulating a subject's immune response in order to improve innate health maintenance capabilities. It has been recognized that there is a relationship between electromagnetic fields and biological functions such as immunity. For example, according to US Pat. No. 4,670,386, the expression of a strong antigenic tumor-specific antigen is induced by exposure of tumor cells to radio frequency electromagnetic radiation, which is a bipolar molecule of cancer cells. Produces a sharp, alternating alternation of polarity in the component. However, one of the drawbacks of very high frequency radiation is that it causes unnecessary heating of biological tissue that can be damaging.

  The relationship between electromagnetic radiation and immunity is also shown in U.S. Pat.No. 6,038,478, where electrodes are applied at desired locations and desired in the body by stimulating the tissue with sufficient current to attract lymphocytes. It is disclosed that lymphocytes can be attracted to the position. In US 2002/0072646, a low energy alternating magnetic field was used to induce an immune response. Similarly, according to WO 02/062418, it is disclosed that the immune function is enhanced by exposing a subject to a magnetic field or a very high frequency electromagnetic field whose angle is adjusted.

  Biological systems often involve electrical activity, as described in chapter 50 of The Biomedical Engineering Handbook (CRC Press 1995). This activity can be a constant DC electric field, a charged particle or a constant flow of current, or a time-varying electric field or time associated with some time-dependent or biochemical phenomenon. There is a possibility of current variation. Bioelectric phenomena are related to the distribution of ions or charged molecules in biological structures, as well as to changes in this distribution caused by specific processes.

  Electromagnetic bioimpedance measurements have been used for diagnostic purposes. For example, according to WO 01/076475, an electric field eddy current is induced in a biological tissue using an alternating magnetic field. An oscillating circuit was used to create a current in the coil adjacent to the target tissue. Since the amplitude of the composite voltage is proportional to tissue conductivity, changes in bioimpedance are used to find tissue changes associated with tumors (such as prostate tumors). Another tissue impedance measuring device for identifying tissue types is disclosed in WO 01/67098. The disclosures of both of these PCT published patents (WO 01/076475 and WO 01/67098) are incorporated herein by reference.

  The effects of electromagnetic fields on biological organisms and their cellular components have long been evaluated. However, it is difficult to take advantage of this interrelationship, and the immune system needs to be more effectively coupled with an external source of regulated electromagnetic radiation.

Protein phosphatases consist of at least two separate independent families: protein serine / threonine phosphatases and protein tyrosine phosphatases (PTPs). Human protein tyrosine phosphatases (human PTPs) are a large and diverse family of proteins present in all eukaryotes. Each PTP consists of at least one conserved domain characterized by an 11 residue sequence motif including cysteine and arginine residues known to be essential for catalysis. The sequence of PTP has no similarity to serine or threonine, acid or alkaline phosphatase. The structural diversity in the PTP family is mainly due to the type of non-catalytic sequence attached to the NH ₂ -or COOH-terminus of the catalytic domain. Many PTPs are associated with intracellular phosphorus metabolism and domains. The diversity of the extracellular region probably reflects the type of ligand that PTPs are exposed to catalyze phosphate transfer.

  PTPs are usually divided into two subgroups. The first subgroup consists of low molecular weight intracellular enzymes with only one phosphatase catalytic conserved domain. All known intracellular PTPs have only one phosphatase catalytic conserved domain. Examples of the first group of PTPs include placental PTP 1B, T cell PTP, rat brain PTP, neuronal phosphatase (STEP), and cytoplasmic phosphatases that have regions of homology with cytoskeletal proteins.

  The second subgroup of PTPs consists of high molecular weight receptor-linked PTPs called R-PTPs. R-PTPs usually contain an intracellular catalytic domain, a single transmembrane domain, and a putative ligand-binding extracellular domain. In contrast to the high homology of the intracellular catalytic domain of R-PTPs, the structure and size of the putative ligand-bound extracellular “receptor” domain of R-PTPs varies greatly. All R-PTPs have two tandem overlapping phosphatase catalytic homology domains, with the exception of the famous R-PTP called HPTPβ, which has only one phosphatase catalytic domain (Tsai et al., J. Biol. Chem. 266 (16): 10534-10543 (1991)).

  An example of R-PTPs is leukocyte common antigen (LCA) (Ralph, S. J., EMBO J. 6: 1251-1257 (1987)). LCA is a family of high molecular weight glycoproteins that are expressed on the surface of all leukocytes and their hematopoietic stem cells. Very high similarity has been found in the sequences of LCAs from various species (Charbonneau et al., Proc. Natl. Acad. Sci. USA 85: 7182-7186 (1988)). LCA is described in the literature as T200, B220 for B cell type, mouse allotype marker Ly-5, and more recently CD45 (Cobbold et al., Leucocyte Typing III, ed. AJ McMichael et al., Pp. 788-803 ( 1987)) is called by different names. CD45 is thought to play an important role in T cell activation. These studies are described in Weiss A., Ann. Rev. Genet. 25: 487-510 (1991).

  Another example of R-PTPs is leukocyte common antigen-related molecule (LAR) (Streuli et al., J. Exp. Med. 168: 1523-1530 (1988)). In addition, published application WO 92/01050 discloses human R-PTP-α, R-PTP-β, R-PTP-γ, and conservation of these three R-PTPs and other members of this protein family. The nature of structural homology found between domains has been reported.

  Extracellular PTPs are related to surface recognition molecules and adhesion molecules for leukocyte cell surface recognition. PTPs are not only associated with human cells, but are also present in prokaryotes and viruses and bacteria. In the pathogen Yersinia, Yop2b tyrosine-specific PTP, a pathogenic factor for glandular plague, is an essential toxicity determinant.

  Many studies have shown the importance of PTPs in physiological processes. Phenotypic abnormalities and hyperproliferative responses of T and B lymphocytes, granulocytes and macrophages are considered important issues in the progression of cancer and autoimmune diseases.

ここで、式中でArは、一つ以上の電子求引基で置換されたアリール基を含み、
XはNH、CH₂、O、―、SO₂、またはB、
Qは-(CH=CH)_m-CH-、
YとY'は独立にN、CHおよびBから選択され、
nは0または1、
mは0から6、
R¹はH、低級アルキル基、随意に置換されたフェニル基または以下の構造で、

ここで式中のR⁴は随意に置換されたフェニル基であり、
R²は随意に置換された低級アルキル基、アラルキル基、チオアルキル基、アミドアルキル基、アミノ基、アミノアルキル基、ニトロ基、アジド基であり、Zと共に随意に置換された4-7員環の炭素環または複素環を形成するか、あるいはR⁴と共に随意に置換されたフェニル基を形成し、
R³はR²と共に随意に置換されたフェニル基を形成するか、または随意に置換されたフェニル基であり、
ZはH、低級アルキル基、置換されたアルキル基、チオアルキル基、二トリル基、アミノ基であるか、またはR²と共に随意に置換された4-7員環の炭素環もしくは複素環を形成する。 Disclosed herein are compounds, compositions, and methods of using such compounds and compositions for modulating, particularly promoting immune responses, which are useful, for example, in the treatment of cancer. In some embodiments, the compound has the following structure:

Here, Ar in the formula includes an aryl group substituted with one or more electron withdrawing groups,
X is NH, CH ₂ , O, —, SO ₂ , or B,
Q is-(CH = CH) _m -CH-,
Y and Y ′ are independently selected from N, CH and B;
n is 0 or 1,
m is 0 to 6,
R ¹ is H, a lower alkyl group, an optionally substituted phenyl group or the following structure:

Where R ⁴ is an optionally substituted phenyl group,
R ² is an optionally substituted lower alkyl group, an aralkyl group, a thioalkyl group, an amidoalkyl group, an amino group, an aminoalkyl group, a nitro group, an azido group, and a 4- to 7-membered ring optionally substituted with Z Form a carbocyclic or heterocyclic ring or form an optionally substituted phenyl group with R ⁴ ;
R ³ together with R ² forms an optionally substituted phenyl group or is an optionally substituted phenyl group;
Z is H, a lower alkyl group, a substituted alkyl group, a thioalkyl group, a nitrile group, an amino group, or an optionally substituted 4-7 membered carbocyclic or heterocyclic ring with R ² .

  In some embodiments, the compound is derivatized with a biomolecule (such as a sugar, amino acid, peptide, fatty acid, nucleoside, or nucleic acid). In some embodiments, such derivatization is used to increase the solubility of the compound and / or to direct the compound to a particular tissue, cell, or receptor. In certain embodiments, the optionally substituted phenyl groups listed for the general structural formula above include such biomolecules.

  In certain embodiments, the disclosed compounds are resonance modifying agents that are effective to promote a therapeutic response. Accordingly, what is disclosed herein is a method of promoting a therapeutic response (such as an immune response) by administering a therapeutically effective amount of a resonance modifying compound to a subject in need of treatment (such as an immune stimulus). This resonance modifying compound has a resonating intramolecular dipole moment (or electron density) and can interact with the biological environment. Administration of the compound can take many forms, including but not limited to topical use at the target site, insertion into the skin, insertion into the affected organ, and inhalation. The resonance modifying compound has the ability to promote an immune response characterized in part by infiltration of immune cells (such as lymphocytes) into target sites around the resonance modifying compound. As immune cells enter the target site, the electromagnetic properties of the target site also change, and these electromagnetic changes can be detected to measure the suitability of the subject's immune response (eg, provided by a resonance modifier). By electromagnetic signal). In this way, for example, due to the lack of expected immune cell aggregation, immune response defects are quickly discovered and appropriate therapeutic or preventive actions are taken. One such clinical practice is to expose the resonance-modifying compound to external electromagnetic stimuli that promote the immune response locally at the target site and remotely throughout the body. Examples of such electromagnetic stimulation include current flowing beyond the potential in the compound, an induced magnetic field, or radiant energy (laser energy, etc.) applied to the compound.

  Particular examples of resonance modifying compounds include aryl compounds, particularly aryl compounds having an aryl group substituted with an electron withdrawing group. In some embodiments, the resonance modifying compound has the general structure described above.

式中のR⁷とR⁸は、独立に、-OR⁵、-SR⁵、-SO₂N(R⁵)₂、-SO₂CN、-SO₂R⁶、-SO₂N₃、N₃、-CN、-N(R⁵)C(O)R⁶、-R⁶であり、
各R⁵は独立にH、低級アルキル基、糖、脂肪酸、アミノ酸、ペプチド、ヌクレオシド、または核酸であり、R⁶は低級アルキル基またはC-グリコシドで、Ar、X、およびYは上記の通りである。 In certain embodiments, disclosed compounds include compounds of the structural formula above (where n is 0), such as, for example,

R ⁷ and R ⁸ in the formula are independently -OR ⁵ , -SR ⁵ , -SO ₂ N (R ⁵ ) ₂ , -SO ₂ CN, -SO ₂ R ⁶ , -SO ₂ N ₃ , N ₃ , -CN, -N (R ⁵ ) C (O) R ⁶ , -R ⁶ ,
Each R ⁵ is independently H, a lower alkyl group, a sugar, a fatty acid, an amino acid, a peptide, a nucleoside, or a nucleic acid, R ⁶ is a lower alkyl group or a C-glycoside, and Ar, X, and Y are as described above is there.

In another embodiment, X represents a chemical bond. Examples of structures in which X represents a chemical bond include the structures shown below.

式中のAr、Y、R⁷およびR⁸は上記の通りである。 An example of an embodiment of such a compound is represented by the following structural formula when n = 0:

Ar, Y, R ⁷ and R ⁸ in the formula are as described above.

式中のGは-CH₂-、-C(O)-、-N-、―、-O-、または-S-をあらわし、nは0または1であり、Y'、Q、Y、X、Ar、R⁷およびR⁸は上記の通りである。 In some embodiments, the compound has the following structure:

G in the formula represents —CH ₂ —, —C (O) —, —N—, —, —O—, or —S—, n is 0 or 1, and Y ′, Q, Y, X , Ar, R ⁷ and R ⁸ are as described above.

式中のR⁵はH、-OR⁶、-SR⁶、-SO₂N(R⁶)₂、-SO₂CN、-SO₂R⁷、-SO₂N₃、N₃、-CN、-N(R⁶)C(O)R⁷、または-R⁷であり、
R¹とR⁶は独立にHまたは低級アルキル基で、
R²とR⁷は独立に低級アルキル基で、Arは上記の通りアリール基である。 Other compounds disclosed herein include those having the following structure:

R ⁵ in the formula is H, -OR ⁶ , -SR ⁶ , -SO ₂ N (R ⁶ ) ₂ , -SO ₂ CN, -SO ₂ R ⁷ , -SO ₂ N ₃ , N ₃ , -CN,- N (R ⁶ ) C (O) R ⁷ , or -R ⁷ ,
R ¹ and R ⁶ are independently H or a lower alkyl group,
R ² and R ⁷ are independently a lower alkyl group, and Ar is an aryl group as described above.

または

または

ここで、式中のR¹は水素、ヒドロキシル基、ヒドロキシフェニル基（2-ヒドロキシフェニル基もしくは4-ヒドロキシフェニル基など）、酢酸塩、リン酸塩、アジド基、二トリル基、アミノ基、ジメチルアミノ基、硫酸塩、メチルスルホン酸塩、リン酸塩、コハク酸塩であり、
R²は非置換の(C₆H₅)か、または置換されたフェニル基（C₆H₄OH、C₆H₄N₃、C₆H₄CN、4-HO-C₆H₄-C₆H₄、C₆H₄OPO₂OH、C₆H₄OSO₂H、C₆H₄NH₂、C₆H₄NHMe₂、C₆H₄OSO₂Me、C₆H₄OCO(CH₂)_xCO₂H、またはC₆H₅Clなど）であり、
ArはC₆H₃-2,4(NO₂)₂、C₆H₄-4(NO₂)、C₆H₄-3(NO₂)、またはC₆H₃-2,4(NO₂)₂などのニトロフェニル基であり、
R³は-O-、-S-、-CH₂-、-N-、―、-CHA-、および-CHOA-であり、ここでAはアリール基、エステル、アミド基、脂質、糖、またはペプチドであり、
YはH、(CH)_xCH₃（x = 0-12）、-S-CH₃、二トリル基、アミノ基、ニトロ基、アジド基、コハク酸塩、またはアミド基であり、
ZはH、(CH)_xCH₃（x = 0-12）、-S-CH₃、二トリル基、アミノ基、ニトロ基、アジド基、コハク酸塩、またはアミド基である。 Particular embodiments of the compounds disclosed herein include aryl hydrazones. Other examples include aryl nitrohydrazones, such as phenyl hydrazones, such as polyaryl mononitro- or dinitrophenyl hydrazones, for example the structures:

Or

Or

Here, R ¹ in the formula is hydrogen, hydroxyl group, hydroxyphenyl group (such as 2-hydroxyphenyl group or 4-hydroxyphenyl group), acetate salt, phosphate salt, azide group, nitrile group, amino group, dimethyl group Amino group, sulfate, methyl sulfonate, phosphate, succinate,
R ² is unsubstituted (C ₆ H ₅ ) or a substituted phenyl group (C ₆ H ₄ OH, C ₆ H ₄ N ₃ , C ₆ H ₄ CN, 4-HO-C ₆ H ₄ -C ₆ H ₄ , C ₆ H ₄ OPO ₂ OH, C ₆ H ₄ OSO ₂ H, C ₆ H ₄ NH ₂ , C ₆ H ₄ NHMe ₂ , C ₆ H ₄ OSO ₂ Me, C ₆ H ₄ OCO (CH ₂ ) _x CO ₂ H, or C ₆ H ₅ Cl),
Ar is C ₆ H ₃ -2,4 (NO ₂ ) ₂ , C ₆ H ₄ -4 (NO ₂ ), C ₆ H ₄ -3 (NO ₂ ), or C ₆ H ₃ -2,4 (NO ₂ ) A nitrophenyl group such as ₂ ,
R ³ is —O—, —S—, —CH ₂ —, —N—, —, —CHA—, and —CHOA—, where A is an aryl group, ester, amide group, lipid, sugar, or A peptide,
Y is H, (CH) _x CH ₃ (x = 0-12), -S-CH ₃ , a nitrile group, an amino group, a nitro group, an azide group, a succinate, or an amide group,
Z is _{H, (CH) x CH 3} (x = 0-12), - S-CH 3, nitrile group, an amino group, a nitro group, an azido group, succinate or an amide group.

  In certain embodiments, these compounds are believed to have resonant intramolecular dipole motion and capable of electrostatic interaction with the biological environment, and in certain embodiments, the extracellular catalytic receptor for R-PTP. Interacts with (such as those found on R-PTP CD45 + PTP subtypes) or generally interacts with lymphocytes. This interaction attracts lymphocytes (such as T lymphocytes) to the target site to which the compound has been applied. Resonance modifying compounds can participate in non-covalent bonds, but usually do not have sufficient chemical reactivity and do not cause local chemical reactions. Nonetheless, the compound appears to activate immune function, for example, by recruiting lymphocytes (eg, T lymphocytes such as CD45 + T lymphocytes, such as CD45RO + or CD45RA + lymphocytes) from the lymphatic network . It also concentrates immune activity at the target site of the body exposed to the compound. Resonance-modifying compounds can also promote the immune system at sites away from the target site (eg, in the spleen, the immune system is promoted by interaction with a distributed network of immune cells such as found in the lymphatic system) ). This mobilization of immune responses can be used to treat infections or tumors.

The disclosed compounds can activate immune system activity away from the site of use of the compound on the body, so that there is substantially no cytotoxicity or other pharmacological action (such as estrogen or antiestrogenic activity) in the target cell itself. In the absence, the compound can act as an immunostimulatory agent.
Since the compound activates an immune response that acts on the target cell, the body's natural defenses are elicited in order to indirectly obtain the desired therapeutic effect. Thus, the drug can remain on the surface of the body (eg, on the skin) and, in order to come into contact with target cells (such as atypical cells, neoplastic cells, or virus-infected cells), it can exert remote immunity without penetrating the skin. Can have. Drugs can also be dispensed in new dosage forms such as tablets or crystals, which provide long-term contact with the environment where immune stimulation is required without necessarily bringing the drug into contact with the cells that are the target of the therapeutic action can do. Other such dosage forms include containers (such as hermetic bandages or impermeable pouches) that are affixed to the skin without direct contact with the subject's body or in the skin. Exerts remote immunoreactive properties without penetrating or through the skin. Alternatively, a remote immune activity characteristic is exhibited by introducing a dosage form into the skin, which is a site that exhibits an effect, in the form of a solid (including crystals). Dosage forms can also include tablets or suppositories, which are introduced into body cavities other than blood vessels (such as the vagina, anus or peritoneum) and fluids that come into contact with the diseased tissue (such as the vaginal neck, anal epithelium, or peritoneal metastases) Dissolve in

  Certain embodiments of the disclosed compounds are cytotoxic. Typical cytotoxic compounds become nuclei or self-assemble in specific tissues, cells or receptors (eg, target cells). In certain embodiments, such compounds are selectively cytotoxic to specific cell types. In certain embodiments, self-aggregation targeted on tumor cells can cause a local increase in drug activity.

  In certain embodiments, the compound is administered by applying to the subject's skin (eg, topically applied to the skin surface with a gel, or otherwise, eg, a crystal or pellet of the compound). Introduce into the skin by intradermal placement). However, compounds can also be administered by introduction into the body (eg, diseased organs or tumors such as malignant or metastatic lesions), thus promoting local immune responses and mobilizing lymphocytes from the lymphatic system And induces an immune response in the target organ or tumor. The compounds can also be administered in aerosol formulations for tracheobronchial or oropharyngeal administration. In certain embodiments, the compound is applied locally to or near a metastatic lesion or epithelial lesion (such as a breast wall breast cancer lesion or cervical epithelial cancer). The compound is applied to the surface of the subject's epithelium or epidermis at a site that is therapeutically close enough to the lesion to promote an immune response at the target site. In the case of superficial lesions (such as breast wall recurrence of breast cancer or cancer of the cervical or anal epithelium), the compound is applied directly to the lesion.

  In other embodiments, the compound is not applied to skin lesions (such as primary tumors or skin metastases), but instead is placed on the body surface overlying the underlying tumor in the body that the compound does not contact. Thereby, the tumor and the compound are separated by a shell structure that the compound cannot penetrate. Thus, the compound is subject to antitumor effects on neoplastic cells deeper in the body (such as metastatic cells in the lymphatic system or tumors in organs and structures such as the uterus, breast, anus, or prostate). Can be applied on the skin (on) or over the skin.

  The disclosed compounds can be used to treat lesions on the internal or external surface of the body. Such lesions can include anogenital lesions such as genital or anal lesions, such as vagina and malignant tumors (such as primary malignant tumors). In certain disclosed embodiments, the lesion is an internal injury, such as a genital wart of a female genital organ (such as a cervical wart). In certain embodiments, the lesion is a precancerous lesion. Indeed, in some embodiments, the lesion is a papillomavirus related lesion, such as an HPV-induced lesion. In yet other examples, the lesion is a non-sexually ill heel, such as a plantar heel, a filamentous heel, a flat heel or a seborrheic heel. In certain embodiments, the disclosed compounds can be utilized to exert a prophylactic effect, such as by treatment of precancerous lesions, as well as prevention or delay of tissue neoplasia. The compounds are particularly suitable for the treatment of genital lesions resulting from a variety of lesions, including vaginal cancer and genital cancer (such as primary penile cancer or vulvar cancer).

  In certain examples, the compound is applied topically to a surface lesion on the body or introduced into the skin (eg, by injection or placement of a pellet into the lesion). Alternatively, the drug is placed in a pad and applied to the lesion, and the pad is left on the lesion for a long time (more than a day or more than a week).

  In other examples, the lesion is a skin lesion resulting from infection or inflammation. The lesion may be a virus-induced lesion, such as a sputum such as a papillomavirus-induced sputum. Alternatively, the lesion to be treated is a skin infection of bacteria (eg, staphylococci) or viruses (eg, herpesvirus), or skin neoplasms (eg, squamous or basal cell epithelial malignancies, or primary melanoma, etc. Skin cancer).

  A compound's immunostimulatory activity, or PTP activation ability, or immunomodulating activity can cause a compound to increase in an electromagnetic field (such as a time-varying electric field, a time-varying magnetic field, and / or a radiating electromagnetic field), such as a resonance modification of the compound It can be improved by exposing to. For example, such a field can be achieved by placing a magnetic probe in the vicinity of a compound administered to a subject, or by applying a current that flows through the compound between two electrodes, or irradiating the compound with laser energy. It can be guided by a laser. In certain embodiments, the activity of the immune system by applying the compound to a target site on the skin using convenient patches or pellets and by inducing an electromagnetic field in a manner that increases the resonance modification of the compound. Is conveniently up-regulated. In this way, the resonance modifier at the skin surface serves as a convenient link between the externally applied electromagnetic field and the immune system, thereby increasing the immune function particularly locally at the site of the resonance modifier. . However, it is considered that the immune function can be enhanced systemically away from the resonance modifying compound.

  The ability to interact with PTPs (such as components of the immune system) allows compounds to be used to monitor immune function. It has been observed that by applying a compound to a target site (eg, in a patch and applied to the skin), PTP expressing cells such as cellular components of the immune system are mobilized. Immune cell attraction is probably caused by electrostatic interactions with dendritic cells and other early lymphoid cells, and attraction of those cells. These cells aggregate near the resonance modifier and change the electromagnetic properties (such as bioimpedance) of the target site where they aggregate. The changed electromagnetic characteristics can be detected even when there is no inter-electrode applied potential difference, for example, as a change in the amplitude of the potential difference detectable across the target site. In certain embodiments, the electrode is placed in contact with the compound, and an intrinsically variable voltage is created over time by resonant modification of the compound. The amplitude of these variable potential differences is used for monitoring immune function. For example, if the voltage amplitude between the electrodes is reduced compared to the normal control, it means that the immune function is reduced.

  Waveform amplitude (or other electromagnetic properties) may be monitored to determine a subject's immune system response to the compound. An increase in voltage amplitude over time means that an immune response is taking place (as well as immune effect cells such as lymphocytes have migrated to the target site). If the increase in amplitude is below a predetermined threshold (such as at least 10%, 25%, or 50% expected increase), further diagnostic action or treatment to determine or correct the cause of reduced immunity Action is performed. For example, rigorous investigations are undertaken for infectious, toxic or neoplastic causes in a reduced immune response. Alternatively, the immune response can be upregulated by exposing the resonance modifying compound to an electromagnetic field that causes an increase in the resonance modification of the compound. Other approaches are also considered to initiate treatment of the subject with an appropriate anti-infectious or anti-tumor chemotherapeutic agent.

  The disclosed agents are particularly effective in treating tumors by administering the agent to a subject (eg, by topical application to a target site on or near skin metastases). Thereafter, an external electromagnetic field is optionally applied to the drug to enhance the resonance modification, thereby enhancing the immune stimulating effect. For example, the external electromagnetic field is created by a magnetic probe that induces a local magnetic field, an induced external current applied between the electrodes, or a laser that promotes the resonance modification of the resonance modifying compound. As a result, recruitment of antigen-presenting cells (such as dendritic cells) and immune effectors (such as T cells) to the target site is increased, which helps direct immune surveillance and immune effect activity to the tumor target . The agents can be used for the treatment of, for example, cervical, anal, or vaginal epithelial malignancies (such as epithelial malignancies for HPV infection, where HPV induces malignant transformation of cells).

  In certain embodiments, the disclosed agents promote an immune response through or against RPTP expressing cells (such as CD45 + cells, such as CD45 + T lymphocytes, or RPTP, or CD45 + expressing infected cells). be able to. Examples of such cells are lymphocytes made in response to virus-infected cells (eg, papilloma viruses such as human papilloma virus (HPV) or cells infected with immunodeficiency virus (such as HIV)).

  In certain methods of monitoring the immune response, the compound is placed in contact with the subject (eg, on or within the subject's skin). In certain embodiments of the method, the compound is a resonance modifier such that the intrinsic resonance modification of the compound results in a characteristic waveform of the compound. Waveforms detected from the compound (such as alternating potential differences or other electromagnetic waves) are monitored to detect waveforms generated from the compound that are associated with altered immune function. In certain embodiments, the waveform is monitored by detecting voltage changes over time in the compound (eg, detecting a waveform with a reduced amplitude over the expected increase seen over time). In some embodiments of the method, once a waveform associated with the altered immune function is detected, a diagnostic or therapeutic action is performed in response to detecting the waveform.

  For example, a medical practice may be characteristic of altered immunity (such as a reduced number or function, or specific cells associated with an immune response such as T lymphocytes), or a cause of altered immunity (changes immune function) , An immunodeficiency virus infection, or the presence of neoplastic diseases such as tumors). Alternatively, treatment that results from the detection of altered immunity, such as administration of an anti-tumor or anti-infection therapy (such as an antibiotic or antiviral agent) and / or modifying the resonance of a compound to enhance immune function Action is brought about. Modifying the resonance of a compound includes applying an induced electromagnetic field to the compound to enhance the resonance modification of the compound, thereby improving immune function. In certain embodiments, the improved immune function improves the recruitment of T lymphocytes (such as CD45 + T lymphocytes) to the site of application of a resonance modifying compound.

  The disclosed method exposes the tumor to a therapeutically effective amount of a resonance-modifying compound and increases the resonance modification of the compound by applying an external electromagnetic field to the compound, thereby increasing the anti-tumor effect of the compound to treat the tumor Further included is a method of treating a tumor by causing. Alternatively, the method may involve administering a therapeutically effective amount of the compound to cause infections (bacterial infections such as plague infections, or viral infections such as HPV and HIV infections, or other such that PTP is expressed by pathogens. It can also be used to treat infectious diseases. In certain embodiments, the therapeutic amount is sufficient to interact with and activate the cellular PTP extracellular receptor. In particular examples of either treatment, the compound is applied to the skin of the subject or to the site of infection or neoplasia. The compound is applied as a topical gel, eg, on the skin of a subject infected with papillomavirus (HPV, etc.) or on the genitourinary or anogenital epithelium (anal, vaginal, or cervical epithelium). Sometimes. In some embodiments, the epithelium is dysplastic or metaplastic epithelium (cervical intraepithelial neoplasia, or high grade neoplasia, or squamous intraepithelial neoplasia (CIN / HSIL), anal intraepithelial neoplasia Organism (AIN / HSIL) or squamous cell carcinoma). In certain examples, the compound is applied as a 0.25% gel that is applied topically to a subject for a period of time sufficient to maintain a therapeutic effect (such as five days or more). In certain treatment methods, the gel is applied daily and / or the effective amount is 2 grams or more of topical gel containing 0.25% or more of the compound.

  Due to the unusual properties of the disclosed compounds, the compounds can be utilized both in vitro and in vivo in methods of enriching dendritic cells and / or lymphocytes. For example, RPTP + cells such as lymphocytes (eg, CD45 + cells such as T lymphocytes) can be enriched from biological tissue (or in culture) by exposing the biological tissue (or cells) to an effective amount of an agent. . In some embodiments, the drug is applied to the subject's skin, and RPTP + cells (such as lymphocytes) are concentrated at and around the site of drug application. However, RPTP + cells (such as lymphocytes and / or dendritic cells) are attracted to any target site in the body (such as breast, colon, or prostate tumors) into which the drug has been introduced. Alternatively, the drug is introduced into tissue culture containing RPTP + cells (such as lymphocytes and / or dendritic cells) and attracts these cells to the drug. In this way, selective enrichment of cell variants is performed. Agents can further be used to increase the enrichment of CD45RO + and CD45RB + cells, for example by inducing the expression of these cell surface markers. In certain embodiments, selective expression of CD45RO + occurs, which is associated with cytotoxicity.

  Further disclosed herein are methods for promoting platelet concentration, eg, in the treatment of thrombocytopenia, eg, by promoting platelet production. One embodiment of this method includes treating a subject with a reduced platelet count using the compounds disclosed herein. In certain embodiments, a subject with thrombocytopenia has a reduced platelet concentration as a result of a neoplastic, autoimmune or infectious disease. In certain examples, the subject suffers from autoimmune thrombocytopenia, or toxic thrombocytopenia (eg, caused by a cytotoxic anticancer drug).

  The foregoing and other materials, features, and advantages of the disclosed compounds, compositions, and methods of use thereof will become more apparent from the following detailed description of some embodiments and are illustrated in the accompanying drawings. To proceed.

[1] Abbreviations
A-007: 4,4'-Dihydroxybenzophenone-2,4-dinitrophenylhydrazone
AMTR: A-007 magnetic transistor resonator
An: Antigen
APC: antigen-presenting cells
BDP-DNP: 2,6-Dibenzylidenecyclohexanone-2,4-dinitrophenylhydrazone
DC: Dendritic cells
DNP: Dinitrophenyl hydrazone
PTP: Protein tyrosine phosphatase [2] Terminology

  Unless otherwise noted, technical terms are used according to conventional usage. Common terms in molecular biology are defined by Benjamin Lewin, Genes V, 1994 (ISBN 0-19-854287-9) published by Oxford University Press, Kendrew et al. (Eds.), The Encyclopedia, published by Blackwell Science Ltd. of Molecular Biology, 1994 (ISBN 0-632-02182-9), Robert A. Meyers (ed.), Molecular Biology and Biotechnology: a Comprehensive Desk Reference, 1995 (ISBN 1-56081-569), published by VCH Publishers, Inc. -8). The variables used in this disclosure are the same as the default variables unless otherwise specified.

  If any of these terms conflict with a document cited by reference, it shall be interpreted as the meaning of this term.

In order to facilitate review of the various embodiments of the present disclosure, the following specific terminology is provided.
Alkyl group: An alkyl group is a branched or unbranched saturated hydrocarbon group having 1 to 24 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t- Examples thereof include butyl group, pentyl group, hexyl group, heptyl group, octyl group, decyl group, tetradecyl group, hexadecyl group, eicosyl group, and tetracosyl group. A “lower alkyl group” is a branched or unbranched saturated hydrocarbon having 1 to 10 carbon atoms.
Aryl group: An aryl group represents any carbon group aromatic group and includes, but is not limited to, a phenyl group, a naptyl group, a fluorenyl group, and the like. The term “aromatic” also includes “heteroaryl groups”. The term heteroaryl group represents an aromatic group having one or more heteroatoms incorporated into the ring of the aromatic group. Examples of heteroatoms include, but are not limited to nitrogen, oxygen, sulfur, and phosphorus. Aryl groups include, but are not limited to, alkyl groups, alkynyl groups, alkenyl groups, aryl groups, halogen atoms, nitro groups, amino groups, ester groups, ketone groups, aldehyde groups, hydroxyl groups, carboxylic acids, or alkoxy groups. It may be substituted with one or more groups that are not, or the aryl group may not be substituted.

走化性：化合物に反応して起こる、生物または単細胞（白血球など）の運動。本明細書で用いるように、走化性は、静電的性質を含む、化合物の任意の物理的性質に反応して起こり得る。
化学療法；化学療法剤：本明細書で用いるように、例えば異常細胞増殖（新生物など）を特徴とする疾病など、疾病の処置に治療有効性を持つ任意の化学薬品。或る実施形態では、化学療法剤は固形腫瘍などの新生物の処置に利用される薬剤である。当業者は、有効な化学療法剤を容易に特定できるだろう（例えば以下を参照。Slapak and Kufe, Principles of Cancer Therapy, Chapter 86 in Harrison's Principles of Internal Medicine, 14th edition、Perry et al., Chemotherapy, Ch. 17 in Abeloff, Clinical Oncology 2^nd ed., （コピーライト）2000 Churchill Livingstone, Inc.、Baltzer L, Berkery R (eds): Oncology Pocket Guide to Chemotherapy, 2nd ed. St. Louis, Mosby-Year Book, 1995、Fischer DS, Knobf MF, Durivage HJ (eds): The Cancer Chemotherapy Handbook, 4th ed. St. Louis, Mosby-Year Book, 1993）。
誘導体：誘導体は、元の化合物から誘導された、あるいは理論的に誘導可能な、化合物または化合物の一部をあらわすために用いられる。
樹状細胞（DC）：樹状細胞は、一次免疫応答に関する主要抗原提示細胞（APCs）である。樹状細胞は、形質細胞様樹状細胞と骨髄樹状細胞を含む。その主な機能は、組織において抗原を特定および処理し、リンパ器官へ移動し、T細胞カスケードを活性化するために抗原情報を提示することである。未熟樹状細胞は骨髄に由来し、未熟細胞として周囲に存在する。
DCsは、未熟な抗原捕獲細胞から、成熟した抗原提示T細胞プライミング細胞へと進化することができ、抗原を免疫原に転換し、免疫反応を開始するために、サイトカイン、ケモカイン、副刺激分子、およびプロテアーゼなどの分子を発現することができる。
ヒドラゾン：構造R₂C=NNR₂を持つ化合物であり、二重結合酸素を=NNR₂で置換している点で、ケトンやアルデヒドとは異なる。ヒドラゾンは、一般にヒドラジンをカルボニル基で縮合することで作られる。アリールヒドラゾンは、一つ以上のR基がアリール基（例えばフェニルヒドラゾンならフェニル基）であるヒドラゾンである。ニトロフェニルヒドラゾンは、一つ以上のNO₂置換基をフェニル環上に持つフェニルヒドラゾンである。
免疫反応：異質（非自己）の物質に対する生物反応。刺激に対する免疫反応は、Bリンパ球またはTリンパ球などの免疫系の細胞によって行われる。或る実施形態では、反応は特定抗原に特異的である（“抗原特異反応”）。
感染物質：対象を感染させ得る物質であり、ウィルス、細菌、および菌類を含むが、これに限定されない。
固有電磁波形：化合物の特性として作られる波形であり、活発誘導された電磁現象（電流、電位、または磁場の意図的な印加など）とは独立している。波形の例は、経時的な交流電圧によって作られる波形、例えば正電位と負電位の間で交流する波形であり、大抵予測可能な形である（例えば正弦波で定められる）。波形は種々の電磁監視装置（化合物と接する二電極間の電位を測定する電圧計など）によって監視できる。
診療行為：診療行為とは、被験者の生理学的状態または医学的状態を発見するか、あるいはその状態に作用するためにとられる行動のことである。診断行為は、例えば臨床試験（血液検査、バイオプシー、画像検査、または身体検査など）によって被験者の状態を検出する。治療行為は、例えば外科手術、薬剤の投与または他の処置、あるいは他の治療手段の実施によって、被験者の状態に影響を及ぼす。
単離した：“単離した”生物学的要素（樹状細胞やリンパ球、またはそれらの細胞の集団など）は、その要素が自然発生する生物の細胞内において、実質的に他の生物学的要素から分離された、あるいは精製されたものである。
免疫不全の臨床検査値：低下した免疫の証拠として、一般に医学的に認められる客観的な検査値であり、特定の研究室の承認基準を下回る白血球数、低下したリンパ球（Tリンパ球など）濃度、または免疫系の任意の細胞の全身活性障害の証拠などである。特定の実施例では、検査値は細胞性免疫または液性免疫の不全、あるいはその両方を示す。
化学反応性欠如：或る共鳴修飾化合物は静電的に活性化しているが、その共鳴安定化のために、生理条件下（体内）においては実質的に非反応性である。そのような化学反応性の欠如の結果、化合物は全身投与（口腔投与または静脈内投与など）の後、実質的に全く変化していない状態で分泌される。この反応性の欠如の例は、例えば体内でA-007のN-メチル化が見られないことによって確かめられる。
白血球：感染性微生物と外来物質に対する体の防御に関連する、“white cells”とも呼ばれる血液中の細胞。白血球は骨髄で作られる。白血球には五つの主要型があり、二つの主要グループに更に細かく分けられる（多形核白血球（好中球、好酸球、好塩基球）および単核白血球（単球およびリンパ球））。感染が見られると、白血球の産生は増加する。
新生物：良性および悪性腫瘍、さらに他の増殖異常を含む、異常細胞増殖。
任意の、または任意に：これらの用語は、後述の事象または状況が起こり得るが、起こらなくてもよいことを意味し、また、前述の事象または状況が起こる場合と、起こらない場合とが含まれた記述であることを意味する。
パピローマウィルス：パピローマウィルスは、正２０面体対称のカプソメアと、約8,000 bpの二本鎖環状DNAゲノムを持つ、小さな非エンベロープウィルスである。全パピローマウィルスは、類似した遺伝子構成を有する。ウィルスゲノムは、ウィルスDNAの複製と細胞形質転換に必要な遺伝子をコードする初期領域と、カプシドタンパク質をコードする後期領域、ならびに、複製開始起点と、多くの転写調節因子と複製調節因子を含む調節領域とに分けられる。
パピローマウィルスは高い種特異性を持つ。ヒトパピローマウィルス（HPV）が、他の種へ自然伝播する例は知られていない。パピローマウィルスは著しい向細胞性も示し、皮膚または粘膜の表面扁平上皮のみに感染し、大部分は良性上皮腫瘍を形成する。特定のウィルス型は、皮膚型または粘膜型のどちらにも嗜好性を有すると思われる。例えば、HPV-11は他の身体部位から皮膚上皮には容易に感染しないが、生殖管または気道のいずれの粘膜上皮も感染させることができる。しかし、パピローマウィルスは浸潤癌の進行につながり得る、細胞増殖と形質転換を誘導する。HPV感染は子宮頸癌と肛門癌の進行に関連している。
生殖器疣は通常HPVに由来する。パピローマウィルスは皮膚と粘膜上に小さな腫瘍（疣）を形成する。生殖器と肛門部位のHPV感染は、陰茎、外陰部、尿道、膣、子宮頸部、および肛門周辺（肛門周囲）上に疣（肛門性器の湿疣）を形成する。50以上の異なるHPVの型が分類されている。6および11を含むいくつかの型は、隆起して面の粗い、容易に見える生殖器疣（特に女性の）に関連している。他の型は、扁平疣に関連する。より重要なことは、いくつかの型は、子宮頸部の前癌変化と悪性変化（異常子宮癌検査）に関連するということである。これらは型16、18、31、39、45、51および52を含む。研究によれば、HPVとヘルペスウィルス両方の存在は、子宮頸癌の進行の良い予測判断材料となることも示されている。
外生殖器上の病変は容易に確認される。陰茎上では、女性生殖器上や両性の肛門周辺に比べ、生殖器疣はより乾燥しやすく貧弱になりやすい。生殖器疣は湿った生殖器部で最も成長し、隆起して面の粗い、肌色の“いぼ状”に見える腫瘍となり、一つずつ、または塊となって形成される。処置されないままでいると、肛門と外陰部周辺の疣は急速に肥大化し、カリフラワー状の外見をとる。女性では、HPVは膣と子宮頸部に侵入する。これらの疣は扁平で、特別な手段なしには容易に見ることができない。HPVは子宮頸部の前癌変化（子宮頸部異形成）を引き起こし得るため、この症状を診断し処置することが重要である。定期子宮癌検査はHPVの発見に重要である。
パピローマウィルス関連部位：病変に関連したHPVの同定など、パピローマウィルス感染の証拠が見られる部位。
薬剤物質または薬剤：被験者に適切に投与されたときに、所望の治療効果または予防効果を引き起こすことができる化合物もしくは組成物。薬剤物質は、化学療法薬剤と抗感染薬剤を含むが、これに限定されない。
薬学的に許容される担体：本発明で有用な薬学的に許容される担体は従来のものである。Remington's Pharmaceutical Sciences, by E. W. Martin, Mack Publishing Co., Easton, PA, 19th Edition (1995)に、本明細書で開示する化合物の薬学的な配送に適した配合と処方設計が述べられている。
一般に、担体の性質は用いられる特定の投与様式に依存する。例えば、非経口製剤は通常、薬学的且つ生理学的に許容される流体（水、生理食塩水、平衡塩類溶液、含水デキストロース、グリセロールなどを賦形剤とする）を含む、注射可能な流体から成る。固体組成（例えば粉末、ピル、錠剤、またはカプセル）として、従来の非毒性固形担体は、例えば医薬品等級のマンニトール、ラクトース、でんぷん、またはステアリン酸マグネシウムを含む可能性がある。生物学的に中性の担体に加えて、投与される薬学的組成物は、湿潤剤または乳化剤、保存料、およびpH緩衝剤などの、少量の非毒性補助剤（例えば酢酸ナトリウムやソルビタンモノラウレート）を含む可能性がある。
精製された：“精製された”という用語は、完全な純度を必要とするわけではなく、むしろ相対語として意図されたものである。従って、例えばリンパ球の精製品とは、その中でリンパ球が体内の自然環境よりも高い濃度で存在するもののことである。
共鳴修飾剤：共鳴分子内双極子運動（もしくは電荷密度）を有し、それによって生物環境と静電的に相互作用が可能な化合物。共鳴修飾剤は、化合物の分子内共鳴によって作られた、発振周波数波の放出によっても特徴づけられる。この共鳴は、免疫の上方制御および／または促進のために、免疫系の細胞（樹状細胞など）との相互作用のための電気的性質を伝えると考えられる。共鳴修飾剤は、免疫細胞を引きつけ、共鳴修飾剤付近の末梢免疫系の標的部位に凝集させることができる。また、いくつかの事例では、循環する免疫細胞（リンパ節または脾臓といった、末梢血およびリンパ系組織の免疫細胞など）に遠隔作用を及ぼす。多くの共鳴修飾剤は結晶構造を持つ。共鳴修飾剤候補の選択のためのアッセイの例は、実施例１４に開示する。
シグマ波：結晶および組織培養から記録されたシグマ波は、心臓および他の体内生物リズムから作られる電気エネルギーの基線以上の波である。従ってシグマ波は、A-007への暴露後、経時的な電圧変化と振幅増加を測定する。
免疫刺激が必要とされる被験者：免疫系の、全般的な刺激または特定の刺激によって利益を得るような症状を持つ被験者。例には、免疫不全の被験者（HIV感染者、あるいは、最近化学療法薬または他の免疫抑制薬を受けた者）、ならびに、免疫反応の促進によって改善され得る症状を持つ者（リンパ腫など、免疫機能を変える病原体または腫瘍に感染した被験者など）が含まれる。或る実施例では、被験者は、例えば感染症または免疫不全のためなど、腫瘍以外の症状のために免疫刺激を必要とする（例えば感染性の、あるいは薬剤由来の免疫不全）。他の実施例では、免疫刺激を必要とする被験者は、低下した免疫機能の臨床検査値を持つ被験者である。
治療有効量：疾病の進行の抑制もしくは予防、または疾病の退行をもたらすのに十分な投与量、あるいは、痛みや腫れなどの、疾病由来の症状を軽減することができる投与量。
血小板減少症：減少した血小板（thrombocyte）数を表す用語。正常血小板値は当業者に知られており、その値は自動化手段で容易に決定できる。正常値は、使用した検出手段次第で実験室によって異なる。しかし、血小板減少症は、血液凝固の低下と病的出血（例えば歯茎、鼻、または内臓からの出血）につながる。
修正された免疫機能に関連する波形：増進した、または低下した免疫機能を持つ被験者に存在するとみとめられる波形。例えば、共鳴修飾化合物を被験者に適用後、経時的に増加する電位差振幅波形など。振幅の増加は、正常な免疫機能の結果であり、免疫細胞（リンパ球など）の動員と、化合物の適用部位への移行を示すものである。しかし、振幅の増加が、健康なときに実施された基線測定において同一被験者で見られるものを下回る場合、あるいは、被験者集団における統計的正常範囲を下回る場合は、修正された（低下した）免疫機能に関連するものとみなすことができる。反対に、正常（特定個人または集団毎に定められる）を超える振幅の増加は、正常以上の免疫機能を示す。
他に記載の無い限り、本明細書で用いる全ての技術用語と科学用語は、本開示が属する技術分野における当業者によって一般に理解されるものと同じ意味を有する。単数形の語“a”、“an”、および“the”は、文脈に明確な他の表示が無い限り、複数形の言及も含む。同様に、文脈に明確な他の表示が無い限り、語“or”（もしくは、または、など）は“and”（および、ならびに、など）を含むものとする。語“含む（comprises）”は、“包含する（includes）”を意味する。加えて、材料、方法、および実施例は、単に実例のためのものであり、限定的なものではない。 Similarly, the term “aralkyl group” refers to an aryl group having an alkyl group attached to the aryl group as described above. An example of an aralkyl group is a benzyl group.
Animal: Living multi-cellular vertebrate organisms, such as mammals and birds. The term mammal includes both human and non-human mammals. Similarly, the term “subject” includes both human and animal subjects.
Antigen: A compound, composition, or substance capable of promoting antibody production or a T cell response in an animal, including a composition that is injected or absorbed into an animal. Antigens react with specific humoral or cellular immunity products, including those induced by heterologous immunogens. The term “antigen” includes all relevant antigenic epitopes.
Cancerous: Represents a malignant tumor or malignant lesion. Examples include primary cancers of the penis, vulva, vagina, cervix, or integument. An example of penile cancer is invasive squamous cell carcinoma. An example of vaginal cancer is squamous cell carcinoma. Any of these tumors can be treated by the methods described herein.
CD marker: A differentiated population (CD) marker that serves as a cell surface marker for different types of lymphocytes. A systematic nomenclature has been developed, and a CD is defined by the binding of a specific monoclonal antibody to a specific human leukocyte antigen (HLA). For example, Roitt et al., Immunology (6 ^th Edition), 2001 discloses further information about CD markers. One subset of CD markers is the CD45 marker, a leukocyte common antigen (LCA). CD45RA, CD45RB and CD45RO are limited LCAs that are a subset of CD45 + cells and are further described in Roitt, et al., Appendix 2.
Some characteristics of these LCAs are shown in the following table.

Chemotaxis: Movement of an organism or single cell (such as a white blood cell) that occurs in response to a compound. As used herein, chemotaxis can occur in response to any physical property of the compound, including electrostatic properties.
Chemotherapy; Chemotherapeutic agent: As used herein, any chemical that has therapeutic efficacy in the treatment of a disease, such as a disease characterized by abnormal cell proliferation (such as a neoplasm). In some embodiments, the chemotherapeutic agent is an agent utilized in the treatment of neoplasms such as solid tumors. One skilled in the art can readily identify effective chemotherapeutic agents (see, eg, Slapak and Kufe, Principles of Cancer Therapy, Chapter 86 in Harrison's Principles of Internal Medicine, 14th edition, Perry et al., Chemotherapy, .. Ch 17 in Abeloff, Clinical Oncology 2 nd ed, ( copyright) 2000 Churchill Livingstone, Inc., Baltzer L, Berkery R (eds):. Oncology Pocket Guide to Chemotherapy, 2nd ed St. Louis, Mosby-Year Book 1995, Fischer DS, Knobf MF, Durivage HJ (eds): The Cancer Chemotherapy Handbook, 4th ed. St. Louis, Mosby-Year Book, 1993).
Derivative: A derivative is used to represent a compound or part of a compound that is derived from the original compound or is theoretically derivable.
Dendritic cells (DC): Dendritic cells are the major antigen presenting cells (APCs) involved in the primary immune response. Dendritic cells include plasmacytoid dendritic cells and bone marrow dendritic cells. Its main function is to identify and process antigens in tissues, move to lymphoid organs, and present antigen information to activate the T cell cascade. Immature dendritic cells are derived from bone marrow and are present in the surroundings as immature cells.
DCs can evolve from immature antigen-capturing cells to mature antigen-presenting T-cell priming cells to convert antigens to immunogens and initiate immune responses, cytokines, chemokines, costimulatory molecules, And can express molecules such as proteases.
Hydrazone: is a compound having the structure R ₂ C = NNR _2, in that to replace the double bond oxygen in = NNR _2, different from the ketone or aldehyde. Hydrazones are generally made by condensing hydrazine with a carbonyl group. An aryl hydrazone is a hydrazone in which one or more R groups are aryl groups (eg, phenyl group for phenyl hydrazone). Nitrophenylhydrazone is a phenylhydrazone having one or more NO ₂ substituents on the phenyl ring.
Immune response: A biological response to a foreign (non-self) substance. The immune response to the stimulus is carried out by cells of the immune system such as B lymphocytes or T lymphocytes. In certain embodiments, the reaction is specific for a particular antigen (“antigen-specific reaction”).
Infectious agent: A substance that can infect a subject, including but not limited to viruses, bacteria, and fungi.
Intrinsic electromagnetic waveform: A waveform created as a property of a compound that is independent of actively induced electromagnetic phenomena (such as the intentional application of current, potential, or magnetic field). An example of a waveform is a waveform created by an alternating voltage over time, such as a waveform that alternates between a positive potential and a negative potential, and is usually in a predictable form (eg, defined by a sine wave). The waveform can be monitored by various electromagnetic monitoring devices (such as a voltmeter that measures the potential between two electrodes in contact with the compound).
Medical practice: A medical practice is an action taken to discover or act upon a subject's physiological or medical condition. The diagnostic action detects the condition of the subject by, for example, a clinical test (such as a blood test, biopsy, imaging test, or physical examination). The therapeutic action affects the condition of the subject, for example, by surgery, administration of drugs or other treatments, or implementation of other therapeutic measures.
Isolated: An “isolated” biological element (such as a dendritic cell or lymphocyte, or a population of those cells) is substantially different from other biology within the cells of the organism in which the element naturally occurs. Separated from the target element or purified.
Laboratory values for immunodeficiency: objective medically accepted objective evidence of reduced immunity, white blood cell count below specific laboratory approval criteria, reduced lymphocytes (such as T lymphocytes) Concentration, or evidence of impaired systemic activity of any cell of the immune system. In particular examples, the test value indicates cellular immunity, humoral immunity deficiency, or both.
Lack of chemical reactivity: Some resonance-modifying compounds are electrostatically activated, but are substantially non-reactive under physiological conditions (in the body) due to their resonance stabilization. As a result of such lack of chemical reactivity, the compound is secreted in a substantially unchanged state after systemic administration (such as buccal or intravenous administration). An example of this lack of reactivity is confirmed, for example, by the absence of N-methylation of A-007 in the body.
White blood cells: Cells in the blood called “white cells” that are associated with the body's defense against infectious microorganisms and foreign substances. White blood cells are made from the bone marrow. There are five major types of leukocytes, which are further subdivided into two major groups (polymorphonuclear leukocytes (neutrophils, eosinophils, basophils) and mononuclear leukocytes (monocytes and lymphocytes)). When infection is seen, white blood cell production increases.
Neoplasm: Abnormal cell growth, including benign and malignant tumors, as well as other growth abnormalities.
Arbitrary or Arbitrary: These terms mean that the events or situations described below can occur but do not have to occur, and include cases where the events or situations described above occur and do not occur It means that it is a description.
Papillomavirus: Papillomavirus is a small non-enveloped virus with an icosahedral symmetrical capsomere and a double-stranded circular DNA genome of approximately 8,000 bp. All papillomaviruses have a similar genetic organization. The viral genome is an early region that encodes genes necessary for viral DNA replication and cell transformation, a late region that encodes a capsid protein, a replication origin, and a number of transcriptional and replication regulatory elements. Divided into areas.
Papillomavirus has high species specificity. There are no known cases where human papillomavirus (HPV) is naturally transmitted to other species. Papillomaviruses are also highly pro-cellular, infecting only the squamous epithelium of the skin or mucous membrane, and most often form benign epithelial tumors. Certain virus types appear to have a preference for either skin or mucosal types. For example, HPV-11 does not readily infect skin epithelium from other body parts, but can infect mucosal epithelium of either the genital tract or respiratory tract. However, papillomavirus induces cell proliferation and transformation that can lead to the progression of invasive cancer. HPV infection is associated with the progression of cervical and anal cancer.
Genital warts usually come from HPV. Papillomaviruses form small tumors (spiders) on the skin and mucous membranes. HPV infections of the genital and anal sites form vagina (anogenital vagina) on the penis, vulva, urethra, vagina, cervix, and around the anus (perianal). Over 50 different HPV types are classified. Several types, including 6 and 11, are associated with genital warts (especially females) that are raised and rough and easily visible. Other types are associated with flat heels. More importantly, some types are associated with precancerous and malignant changes in the cervix (abnormal uterine cancer testing). These include molds 16, 18, 31, 39, 45, 51 and 52. Studies have also shown that the presence of both HPV and herpes virus provides a good predictor of cervical cancer progression.
Lesions on the external genitalia are easily identified. On the penis, genital warts are more prone to dryness and poorness than on female genitals or around the anus of both sexes. Genital folds grow most in the wet genital area and form a bumpy, rough, skin-colored “wart-like” tumor, one by one, or a lump. If left untreated, the anus and the vulva around the vulva rapidly become enlarged and have a cauliflower-like appearance. In women, HPV penetrates the vagina and cervix. These wrinkles are flat and cannot be easily seen without special means. Since HPV can cause precancerous changes in the cervix (cervical dysplasia), it is important to diagnose and treat this condition. Periodic uterine cancer testing is important in detecting HPV.
Papillomavirus-related sites: Sites where there is evidence of papillomavirus infection, such as the identification of HPV associated with the lesion.
Drug substance or drug: A compound or composition capable of causing a desired therapeutic or prophylactic effect when properly administered to a subject. Drug substances include, but are not limited to, chemotherapeutic drugs and anti-infective drugs.
Pharmaceutically acceptable carriers: The pharmaceutically acceptable carriers useful in the present invention are conventional. Remington's Pharmaceutical Sciences, by EW Martin, Mack Publishing Co., Easton, PA, 19th Edition (1995) describes formulations and formulation designs suitable for pharmaceutical delivery of the compounds disclosed herein.
In general, the nature of the carrier will depend on the particular mode of administration being employed. For example, parenteral formulations usually consist of injectable fluids, including pharmaceutically and physiologically acceptable fluids (with water, saline, balanced salt solutions, hydrous dextrose, glycerol, etc. as excipients). . As a solid composition (eg, a powder, pill, tablet, or capsule), conventional non-toxic solid carriers may include, for example, pharmaceutical grade mannitol, lactose, starch, or magnesium stearate. In addition to biologically neutral carriers, the administered pharmaceutical compositions contain small amounts of non-toxic adjuvants such as wetting or emulsifying agents, preservatives, and pH buffering agents such as sodium acetate and sorbitan monolaur. Rate).
Purified: The term “purified” does not require full purity, but rather is intended as a relative term. Thus, for example, a purified lymphocyte product is one in which lymphocytes are present at a higher concentration than the natural environment in the body.
Resonant modifier: A compound that has a resonant intramolecular dipole motion (or charge density) that allows it to interact electrostatically with the biological environment. Resonance modifiers are also characterized by the emission of oscillating frequency waves created by intramolecular resonance of the compound. This resonance is thought to convey electrical properties for interaction with cells of the immune system (such as dendritic cells) for the up-regulation and / or promotion of immunity. Resonance modifiers can attract immune cells and aggregate them at target sites in the peripheral immune system near the resonance modifier. In some cases, it also has a remote effect on circulating immune cells (such as immune cells in peripheral blood and lymphoid tissues, such as lymph nodes or spleen). Many resonance modifiers have a crystal structure. An example of an assay for selection of a resonance modifier candidate is disclosed in Example 14.
Sigma Waves: Sigma waves recorded from crystal and tissue culture are waves above the baseline of electrical energy created from the heart and other body rhythms. Thus, sigma waves measure voltage changes and amplitude increases over time after exposure to A-007.
Subjects in need of immune stimulation: Subjects with symptoms that benefit from general or specific stimulation of the immune system. Examples include immunocompromised subjects (HIV infected or those who have recently received chemotherapeutic drugs or other immunosuppressive drugs), and those with symptoms that can be ameliorated by promoting an immune response (such as lymphoma) Such as a subject infected with a pathogen or tumor that changes function). In certain embodiments, the subject requires immune stimulation due to symptoms other than tumor, such as due to infection or immunodeficiency (eg, infectious or drug-derived immunodeficiency). In other examples, a subject in need of immune stimulation is a subject with a laboratory value for reduced immune function.
Therapeutically effective dose: A dose sufficient to inhibit or prevent progression of the disease, or to cause regression of the disease, or a dose that can alleviate disease-derived symptoms such as pain or swelling.
Thrombocytopenia: A term that refers to a decreased number of thrombocytes. Normal platelet values are known to those skilled in the art and can be readily determined by automated means. The normal value depends on the laboratory depending on the detection means used. However, thrombocytopenia leads to decreased blood clotting and pathological bleeding (eg bleeding from the gums, nose or viscera).
Waveforms associated with modified immune function: Waveforms that appear to be present in subjects with increased or decreased immune function. For example, a potential difference amplitude waveform that increases with time after a resonance modifying compound is applied to a subject. The increase in amplitude is a result of normal immune function and indicates the recruitment of immune cells (such as lymphocytes) and the transfer of compounds to the site of application. However, modified (decreased) immune function if the increase in amplitude is below that seen in the same subject in baseline measurements performed when healthy, or below the statistical normal range in the subject population Can be considered related to Conversely, an increase in amplitude over normal (defined for a particular individual or population) indicates an immune function above normal.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. The singular words “a”, “an”, and “the” include plural references unless the context clearly dictates otherwise. Similarly, unless the context clearly indicates otherwise, the word “or” (or or etc.) shall include “and” (and and etc.). The word “comprises” means “includes”. In addition, the materials, methods, and examples are illustrative only and not limiting.

[3] Description of Some Embodiments Certain methods disclosed herein relate to the use of hydrazone compounds to treat infectious, neoplastic and / or inflammatory diseases. Exemplary methods include convenient and effective treatment for body surface lesions, such as infectious, neoplastic or inflammatory lesions (eg, sputum).

  Without being bound by theory, certain compounds act as resonance modifiers and act as PTPs activators. In some embodiments, the compound acts as an immunostimulant and / or acts as a coupling agent between the immune system and an external monitoring device or modulator. Compounds are thought to interact with PTPs, such as interacting with cellular components of the immune system (such as CD45 + receptor lymphocytes or dendritic cells) to promote immune cell maturation, and other cells of the immune system It may be possible to mobilize components to an immune response. Since compounds can attract immune cells to the vicinity of the modulator, they can direct the immune response to target sites in the body (such as tumors) where an immune response is required for treatment of local symptoms. However, in certain embodiments, the compound can elicit an immune response at a distant site (such as a distant lymphoid tissue or metastatic lesion). Thus, the compounds can provide new contacts with the immune system, collect information about immune status, and perform medical practice (including manipulation of immune function).

  The resonance structure represents the synthetic electronic structure of the compound, where the electron density is delocalized. Multiple alternative formal structures are referred to as resonance structures, and a molecule can be described as a resonance hybrid of these structures. The conjugated aromatic compounds disclosed herein generally comprise a plurality of alternative formal structures with delocalized electron densities.

であり、式中のR¹はOHで、R²はC₆H₄OHで、XはC₆H₃-2,4(NO₂)₂である。共鳴修飾化合物の多くの他の例も本明細書で開示し、その中のいくつかは添付の図１２、１６、および１７に示した。これらは、ホルミルおよびアセチルバルビツル酸フェニルヒドラゾンの類似体を含む。他の例は、2,6-ジベンジリデンシクロヘキサノン-2,4-ジニトロフェニルヒドラゾンで、

式中のXはニトロフェニル（C₆H₃-2,4(NO₂)₂）で、R¹はHで、R²は非置換のフェニル基で、R³はCH₂である。 A specific example of a resonance modifier disclosed herein is 2,4-dinitrophenylhydrazone (referred to as A-007), where the compound is

In which R ¹ is OH, R ² is C ₆ H ₄ OH, and X is C ₆ H ₃ -2,4 (NO ₂ ) ₂ . Many other examples of resonance modifying compounds are also disclosed herein, some of which are shown in the accompanying FIGS. These include analogs of formyl and phenylhydrazone acetylbarbiturate. Another example is 2,6-dibenzylidenecyclohexanone-2,4-dinitrophenylhydrazone,

X in the formula is nitrophenyl (C ₆ H ₃ -2,4 (NO ₂ ) ₂ ), R ¹ is H, R ² is an unsubstituted phenyl group, and R ³ is CH ₂ .

  The resonance modifying compounds disclosed herein include 4,4′-dihydroxybenzophenone-2,4-dinitrophenyl hydrazone (A-007), which generally has an affinity for cell membrane receptors, and therefore has detection agents and It is particularly suitable for working as an immunomodulator. An example of this affinity for the RPTP + cell membrane receptor is the interaction between A-007 and the CD45 + T lymphocyte surface receptor and is shown in FIG. These interactions are thought to induce maturation of interacting cells and promote immune responses.

などであり、式中のR¹は水素、ヒドロキシル基、2-ヒドロキシフェニル基もしくは4-ヒドロキシフェニル基、酢酸塩、リン酸塩、アジド基、二トリル基、アミノ基、ジメチルアミノ基、硫酸塩、メチル硫酸塩、リン酸塩、ニトロソ基、琥珀酸塩、もしくは、水素結合が可能な他の水溶性求電子基であり、R²はC₆H₅、C₆H₄OH、C₆H₄N₃、C₆H₄CN、4-HO-C₆H₄-C₆H₄、C₆H₄OPO₂OH、C₆H₄OSO₂H、C₆H₄NH₂、C₆H₄NHMe₂、C₆H₄OSO₂Me、C₆H₄OCO(CH₂)_xCO₂H、またはC₆H₅Clであり、ArはC₆H₃-2,4(NO₂)₂、C₆H₄-4(NO₂)、C₆H₄-3(NO₂)、またはC₆H₃-2,4(NO₂)₂である。 Example 1 Resonant Modification with Phenylhydrazone This example discloses a particularly suitable conjugated aromatic compound having both modification and detection properties. Some such compounds include polyarylmononitro- or dinitrophenylhydrazones, for example

R ¹ in the formula is hydrogen, hydroxyl group, 2-hydroxyphenyl group or 4-hydroxyphenyl group, acetate, phosphate, azide group, nitryl group, amino group, dimethylamino group, sulfate , Methyl sulfate, phosphate, nitroso group, oxalate, or other water-soluble electrophilic groups capable of hydrogen bonding, R ² is C ₆ H ₅ , C ₆ H ₄ OH, C ₆ H ₄ N ₃ , C ₆ H ₄ CN, 4-HO-C ₆ H ₄ -C ₆ H ₄ , C ₆ H ₄ OPO ₂ OH, C ₆ H ₄ OSO ₂ H, C ₆ H ₄ NH ₂ , C ₆ H ₄ NHMe ₂ , C ₆ H ₄ OSO ₂ Me, C ₆ H ₄ OCO (CH ₂ ) _x CO ₂ H, or C ₆ H ₅ Cl, Ar is C ₆ H ₃ -2,4 (NO ₂ ) ₂ C ₆ H ₄ -4 (NO ₂ ), C ₆ H ₄ -3 (NO ₂ ), or C ₆ H ₃ -2,4 (NO ₂ ) ₂ .

As shown in FIG. 3, these chemical structures can not only resonate between multiple dipole structures in places with many / few conjugated electrons, thereby allowing intermolecular hydrogen bond migration, Nucleophilic / electrophilic attraction with a complementary site is possible so that a compound can be present at a certain moment. As shown in FIG. 1, these resonant small dipoles are, for example, -SH, -COOH, -NH _2- present in amino acids (cysteine, threonine, arginine, etc.), sugars (mucopolysaccharides), and glycoproteins. CH-NH ₃ ⁺ can be attracted, all of which are essential components of the cellular regulators of the immune system. For example, a simple interaction with 3 or 4 amino acids at the CD45 + RA receptor surface on lymphocytes is thought to induce the expression of modified surface proteins (RO, RB), which are expressed in tissues, organs and The T cell cascade can be initiated with the influx of CD4 + / CD8 + into the circulation. In the “moment in time” during which hydrogen bonding and attraction are regulated via electrical stimulation, the transient electrostatic interactions that are inherently associated with these structures increase, and NMR spectroscopy Can be quantified.

  The presence of multiple negatively charged moieties capable of multiple resonant structures is placed through conjugated and aryl bonds, creating an oscillating frequency wave. Certain resonance modifiers described herein (such as A-007) have a sufficient “electron count” and have excessive electrons for electrostatic interactions. The surplus allows attraction of electrophilic / nucleophilic centers present in proteins and other biomolecules, which must react (via electron transfer and covalent bond formation) for structural stabilization. No. Instead, simple “flirtations” with the environment are thought to induce changes in other multimolecular structures (such as the CD45 + receptor) without changing the structure of the resonance modifier.

In certain disclosed embodiments of resonance modifiers having the above structure, R ¹ is OH, R ² is C ₆ H ₄ OH, and X is C ₆ H ₃ -2,4 (NO ₂ ) ₂ (A-007). This compound has a strong negative charge. Further, as shown in FIG. 3, like the additional compounds disclosed herein, A-007 can be represented by several resonance structures. The typical oscillating frequency wave produced by hydrazone interacts electrostatically with dendritic cells and other early lymphoid cell components to promote immunity. This interaction can be incorporated into one or more Dirac equations to formulate the quantum molecular changes in the skin that accompany the influx of diagnostic lymphocyte components that may be selected.

  Stabilization of certain resonance structures of the disclosed compounds is believed to occur upon binding, resulting in changes in frequency and electrical divergence from the test site where the resonance modifier interacted with lymphocytes. This interaction is generally a non-covalent interaction such as electrostatic interaction. The resonance period is inhibited by this interaction, and such interference is monitored (eg, by detecting voltage waveform changes over time) and used as an indicator of activity. The type of waveform pattern indicates the presence of a developing cellular immune response (such as dendritic cell and lymphocyte infiltration). The lack of expected waveforms, or other detected interference patterns, can also help find abnormal immune responses that require further investigation and / or treatment. Thus, the resonance modifier acts as a coupling agent between the immune activity and the external monitoring device.

  Resonance modifiers not only detect cellular immune activity, but also bind the immune system to external physical modulators that can be used to alter this activity. For example, a magnetic resonator probe is used to apply a magnetic field to the skin surface, for example, by applying the probe to or near the skin surface. The magnetic probe activates the resonance modifier, stimulates the outflow of resonance energy, and promotes tissue T cell contact. Thus, if an individual is unable to induce immune modulation via unstimulated T cell-resonant modifier interactions, additional polarity formation provided by an external programmed magnetic field is added to the immune response. Additional stimuli can be provided. Use of such an external magnetic field together with A-007, which is a resonance modifier, is called AMTR (A-007 magnetic transistor resonator).

  Accordingly, some of the disclosed methods relate to the use of various resonance modifiers, such as hydrazones, that can function as immunochemical quantum chemical modifiers / detectors / transmitters. These agents are therefore useful for recording and treating changes in epithelial surfaces (such as skin) associated with lymphatic circuits via biological networks. Peripheral regulation of lymphocytes and progenitor cells in the skin is an early warning network of systemic lymphocytes, and the biological system discovers foreign changes and foreign bodies (viruses, bacteria, chemicals, cancer cells, etc.) It is a natural mechanism. Cancer and exogenous chemical / biological material entry can cause electrical and sudden behavioral changes that require monitoring / modification to protect health. This ability to monitor the status of peripheral regulation therefore provides important new medical diagnostic and therapeutic tools.

[Example 2] Antitumor and immunomodulatory properties of 4,4'-dihydroxybenzophenone-2,4-dinitrophenylhydrazone (A-007) This example includes 4,4'-dihydroxybenzophenone-2,4 -The disclosed compounds, including dinitrophenylhydrazone, show significant anti-cancer activity in anti-cancer tests (IND 47,470). In particular, in 53 people treated with topical A-007 (as a 0.25% gel), 37% objective remission with 10 complete remissions was observed.

  Certain disclosed compounds do not act through cytotoxic mechanisms. This is evidenced by the substantial absence of local or systemic toxicity from, for example, A-007. Histochemical analysis from human skin biopsies treated locally with A-007 confirmed that infiltrates of various T-lymphocytes (CD4 +, CD3 +, CD8 + and CD45 +) increased during treatment. . An increase in skin infiltrate of CD11c + dendritic cells was also observed at the treatment site. Previously, immunohistochemical (IHC) studies have shown that immunomodulation occurs in vitro and in vivo following exposure to A-007. Accordingly, the compounds disclosed herein can have a therapeutic effect without extensive cytotoxicity, often causing concomitant clinical harm.

According to X-ray crystallography data, A-007 (monoclinic crystal) originally exists as two intrinsic molecules, which differ only in the orientation within the bis-diphenylmethane group. Here the rings are roughly perpendicular to each other as shown in the accompanying figures (and are rotated approximately 90 ° from the ring geometry in each rotamer). Both rotamers show a strong intramolecular hydrogen bond between -NH in the -HN-N = C- moiety and oxygen in the o-nitro group. Thus, A-007 has at least three unique moieties, dihydroxy-diphenylmethane, hydrazone, and dinitrophenyl moieties that are thought to contribute to its overall biological activity. However, despite its high electrophilicity, A-007 is substantially lacking in chemical reactivity under physiological conditions in the human body. For example, the NH of the hydrazone group is normally expected to be methylated or acylated in vivo. However, virtually no such change is seen when A-007 is applied to human subjects. In practice, the drug is excreted substantially unchanged. Furthermore, A-007 is very insoluble in body fluids and is not widely absorbed into blood. No A-007 was detected in blood from any of the subjects treated with topical A-007. The plasma analysis of A-007 demonstrates its stability.

Example 3 Effect on the Immune System The lymphatic system includes lymphatic vessels that communicate with other structures and organs, including lymphoid tissue in a differentiated form of network connective tissue. Lymphatic vessels include capillary lymphatic vessels that are connected to larger lymphatics. This lymphatic vessel is similar in structure to a vein, but has thinner walls and more valves. Lymph nodes are distributed throughout the body and are most concentrated in the face and neck, pits, chest cavity, intestine, groin, elbow and knee. Shallow lymphatic vessels generally follow veins, while deeper lymphatic vessels generally follow arteries. These lymph vessels serve to network the lymph in the body, but they are also an important decentralized aspect of the immune system and have the function of monitoring and protecting against foreign cells such as microorganisms and cancer cells.

  The lymphatic system includes a number of lymphocyte types. Some of these lymphocytes are T-cells that directly or indirectly destroy foreign cells by releasing cytotoxic substances. Other lymphocytes are B-cells that differentiate into plasma cells that secrete antibodies against the antigen to help remove the antigen. The lymph nodes filter foreign substances carried by the lymph, and the separated foreign substances are then destroyed by phagocytosis. The spleen, thymus and tonsil are other lymphoid organs that make B-cells, T-cells, and other lymphocytes.

  When tumor cells are present in the dermis and epidermis, dendritic cells from the lymphatic system interact with the tumor cells and mobilize other immune effectors (such as T-lymphocytes) from the lymphatic vessels.

  CD45 + surface receptors are present on lymphatic endothelial cells, particularly dendritic cells. Dendritic cells (DC) are antigen presenting cells (APCs) involved in the initiation of immune responses. DCs that act as precursors to the immune system are involved in antigen capture (An) and subsequent transport to T-lymphocyte-rich sites. DCs are present not only in lymphoid organs but also in lymphoid tissues such as peripheral skin tissues. When immature DCs interact with antigens and become activated, mature APCs are capable of specific immune responses. Secondary lymphoid organs such as the skin mobilize both naive T-lymphocytes and An-activated DCs (APCs) to T-cell rich lymphoid areas / networks (eg, nodes). Co-localization of these initial immune responses constitutes cognitive T-cell activation.

  Both cancer and applied chemicals can produce catastrophic reactions in healthy skin, with collective dispersal information for lymphocyte populations. An effective cognitive response requires both DC (APCs) and lymphocyte cytokine effectors. Chemicals and tumor cells are not effective regulators of APCs because they often have limited expression of microhistochemical (MHC) antigens and lack co-promoting molecules. One mechanism for the progression and development of cancer and similar diseases is the lack of MHC antigenic properties, otherwise a sudden response is produced in the lymphatic network. Similarly, life-threatening chemicals and / or biological contacts can induce sudden lymphatic response patterns. Agents disclosed herein include A-007 and are sufficiently charged to act as a hapten / An and / or through electromagnetic field effects (EFE) for modulation or up-regulation of the neoplastic lymphocyte network. It is an organic molecule. The disclosed compounds comprise a new class of regenerating molecules, referred to herein as resonance modifiers. CD45 + receptor upregulation is thought to be one initiation site for A-007-induced immunomodulation observed in cancer patients.

  CD45 + is expressed as a protein tyrosine phosphatase (PTP) on dendritic cells, lymphocytes, monocytes, and leukocytes, as well as some neoplastic cells, and is involved in phosphorylation of tyrosine residues with other members of PTPs To do. Blocking the CD45 + receptor site with an anti-CD45 antibody suppresses T-cell activation and prevents activation of naïve T-cell mitogens (lectins). The CD45 + receptor surface contains arginine, serine / threonine, and cysteine sites that can bind to natural ligands and / or carry natural ligands to the surface of APCs and can also hydrolyze tyrosylphosphatase . Some of the disclosed compounds, including for example A-007, do not suppress or inhibit CD45 +, but bind CD45 + lymphocytes and dendritic cells (against APCs) to Arg, Cys, Ser / Threo, etc. / Up-regulate via non-covalent bonds (as shown in FIG. 1). Another resonance state of A-007 can interact with another immune cell.

  A-007 activated DCs may initiate division events with naïve human peripheral blood mononuclear cells (PBMC) and up-regulate both CD45 + and CD11c + receptors in human peripheral dendritic cells it can. Dendritic cells in cancer tissue are upregulated from CD45RA + to CD45RO + / CD45RB + following topical A-007 exposure during local treatment of skin lesions. Thus, A-007 is not an inhibitor of CD45 + but an upregulator or modifier of the molecular site. These properties allow A-007 crystals or implanted skin pellets to increase both local as well as more distant immune cell populations (such as splenic dendritic cell populations), as well as CD8 + cytotoxic lymphocytes (CTL), etc. The concentration of immune effect cells can be increased. The effects of functional group substitution on the intramolecular / intermolecular hydrogen bond and electrostatic interaction of A-007 are shown below.

  This evidence indicates that A-007 can induce antigen-presenting cell (APC) maturation by interacting peripherally with dendritic cells on the skin and other epithelial surfaces. In addition, one or more cell surface receptors (CD45 +) appear to be upregulated by A-007. This initial interaction is thought to be via dendritic precursors and other lymphocyte precursors, such as antigen presenting cells. Through this chemical interaction, APCs and other early recognition cells can be sensed and modified along with the maturation of lymphocyte recognition cascades (CD4 +, CD8 +, etc.) and related cytokines.

  Some of the special properties of A-007 that make it suitable as an immunomodulator are that the crystals align in a special pattern when deposited on a glass slide between two wires with an electric current introduced. Is indicated by being able to. This crystal configuration is obtained by evaporating an alcohol (5%) solution of A-007 in air on a glass slide between two wires connected to the positive and negative lead wires of a 9 volt battery, respectively. It was. A-007 condenses in a circular shape at the anode, not as a “point” diffused throughout the field. When the deposition of the alcohol solution takes place in a magnetic field, a single line of tightly agglomerated crystals is created in the center of the slide. Therefore, A-007 has electromagnetic properties that interact with the induced electromagnetic field. This feature can also be used as a screening test for other resonance modifiers.

  These special electromagnetic properties of A-007 are further explained by the conductivity of this compound. This was measured from an A-007 crystal fitted with two microelectrodes using the magnification of a dissecting microscope. The electrodes were separated about 1.5-2 mm on the crystal surface and the energy was measured from the contacts. A Fluke ScopeMeter was used to record the resulting frequency (Hz). There was no externally applied current. Thus, the measured frequency represents the natural physical properties of the crystal. These deviations from the reference frequency are thought to be a translational type of energy that accumulates from the environment and is intermittently released. The reference frequency appears to be derived from environmental electromagnetic radiation. Deviations from a reference frequency of 30-40 Hz or higher are created by crystal resonance due to intermittent deviations (indicated by the peak of the frequency) from a reference frequency of approximately 60 Hz. Oscillate the natural frequency of energy (eg, at frequencies above 20 Hz from the reference, eg, between 30-40 Hz) and / or at least once every 5 minutes (eg, once every 3 minutes) This ability is another example of the characteristics of a resonance modifier, which allows the resonance modifier to be screened and selected for further testing.

  When chemical crystals or pressurized pellets are soaked in tissue culture medium with naïve human lymphocytes (obtained from peripheral blood), lymphocytes and dendritic cells aggregate around the crystals or pellets. A-007 crystals (0.5 mg) were placed in RPMI solvent containing 5% bovine serum albumin, antibiotic preservative penicillin / streptomycin, and naive lymphocytes obtained from buffy coats of healthy human blood. It was. Not only did aggregation and increased mitotic activity occur after contact with the crystals of A-007, but also the establishment of activated dendritic cells at the site of the A-007 resonance modifier.

  The ability of an agent to aggregate unsensitized lymphocytes in culture around the test agent determines whether the agent is a suitable resonance modifying agent for use in the methods disclosed herein. Above is another factor to be considered. The ability to induce aggregation within an hour particularly strongly suggests that the compound is suitable for further study. “Aggregation” refers to a substantial increase in the number of lymphocytes, such as an increase that can be seen with a microscope.

  Another special electromagnetic property of resonance modifiers is that the increase in amplitude of electromagnetic waves (such as current) can be measured as mature, aggregated lymphocytes around the crystal.

  To obtain these waveforms, A-007 was condensed onto a sterile glass microscope slide as shown in FIG. Immerse the dried slide in a Petri dish containing RPMI (5% bovine albumin), insert a pair of sterile microelectrodes into the solvent (approximately 5 cm apart), and connect it to the opposite end of the A-007 line. And voltage was recorded. A-007 does not dissolve in the solvent and remains on the slide continuously. Since no induced current was present, the measured current became an intrinsic property of the resonance modifying compound.

  One hour after exposure to A-007, lymphocytes migrated and adhered to the A-007 crystals, and the amplitude of the measured voltage increased over time. Since crystals do not melt, they can continue to be utilized for continued interaction. In vitro, lymphocytes eventually die after about 24 hours due to the lack of cytokines necessary for cell perpetuation.

  Thus, another feature available for the selection of resonance modifiers increases the amplitude of the alternating voltage in the intrinsic current produced by the resonance modifier when the compound is placed in culture containing naive lymphocytes. Is ability.

Example 4 Monitoring Immune Function As a result of its exceptional properties, the compounds described herein (including resonance modifiers) can also be used to monitor electrical activity associated with the immune system. Exemplary compounds can be used as analyzers to measure and control immune properties, which have a linear relationship between T-lymphocytes and frequency response. There is a microbalance due to the fact that the mass sensitivity of a 50 Hz A-007 crystal is approximately 0.057 Hzcm ² ng ^-1 , which is about 50 times higher than the sensitivity of an electron microparticle balance with a sensitivity of 0.1 μg. . Crystals can be used to measure the electromagnetic properties of cellular components. In certain embodiments, a crystal is selected that has a mass sensitivity of at least about 0.01, 0.03, or 0.05 Hz cm ² ng ⁻¹ . Crystals can be combined with quartz crystallites to amplify electrical interactions (piezoelectric resonator effects) and improve transmission. In particular examples, the quartz crystallites are provided in an amount of 10-50% of the total weight of the composition.

  Thus, crystals can easily transform the interaction between a population of RPTP + cells (such as lymphocytes) and the corresponding electrical circuit of the skin tissue, thereby allowing for the presence of hydrazone and other resonance modifiers. Allows a complete description of the amplitude at. Basically, resonance modifiers act as resonators for dipolar motion and electrostatic interactions with cellular components (such as cells of tissues and the peripheral immune system). Resonant modifiers can monitor these interactions for diagnostic purposes and modify them for therapeutic action.

  The availability of resonance modifying compounds for monitoring immune function allows for early diagnosis of a subject's modified ability to recognize biological damage such as toxicants and foreign antigens. For example, subjects exposed to foreign chemicals or biological factors may not perceive exposure and as a result may not respond appropriately. However, if the resonance modifier finds a change in the functioning of the immune system, this modified immune condition acts as a precursor event that warns the subject of exposure to an unknown unknown toxin. Similarly, subjects exposed to environmental stress (such as viruses / bacteria and traumatic events) may have dendritic cells that do not recognize foreign viruses, chemicals, or cancer. Dendritic cell dysfunction can lead to serious invasion of immune defenses and life-threatening through sudden behavioral mechanisms.

  Resonance modifiers may also be used in methods to detect and quantify changes in skin cell geometry associated with normal body immunity and natural surveillance activities. According to animal and human experiments, resonance modifiers such as A-007 are not absorbed from the skin, but are required for natural regulation of foreign body contamination or inflammation, dendritic cells, CD8 + cytotoxic lymphocyte populations, and other It has been shown that it will attract and activate the cell population. If the body does not respond to the presence of the presented detection agent / stimulus, then active health care and treatment continues.

  The monitoring device may take the form of a patch that contacts the subject's skin and retains the resonance modifier. The patch can be periodically connected to a voltmeter, such as a voltmeter that outputs in the form of a tracking waveform of the type obtained from an oscilloscope. Thereafter, the amplitude of the voltage change over time is evaluated and compared to the change observed in healthy subjects to determine how the amplitude has changed. In certain embodiments, the amplitude of the voltage waveform is expected to increase by a predetermined value (eg, at least 10% or 25%) in the presence of the resonance modifier. A change in amplitude below a predetermined value is considered an indicator of pathological damage, such as exposure to a toxin or pathogen. This result suggests the need for more specialized tests such as the discovery of environmental toxicants or pathogens, or imaging tests to detect tumors and tumor progression. Alternatively, an abnormal test can facilitate the initiation of treatment (or more aggressive treatment), such as administration of a chemotherapeutic agent.

  The observed intrinsic amplitude and amplitude changes appear to vary with each resonance modifying compound. Therefore, a specific amplitude change cannot be expressed in all resonance modifying compounds. Nevertheless, recognition of this property of resonance modifiers and the ability to serve as a contact point for immune function is the basis for the disclosed method. Once this property has been confirmed, it is possible to determine the specific intrinsic amplitude produced by each compound and the amplitude change seen in a particular disease state or systemic immunodeficiency.

  In some embodiments, a piezoelectric crystal may be introduced into the resonance modifying compound to provide maximum A-007 transmission through the transducer effect.

式中のArは、一つ以上の電子求引基で置換されたアリール基を含み、
XはNH、CH₂、O、―、SO₂、またはBで、
Qは-(CH=CH)_m-CH-で、
YおよびY'は独立にN、CH、およびBから選択され、
nは0または1で、
mは0から6で、
R¹はH、低級アルキル基、随意に置換されたフェニル基、または

であり、式中のR⁴は随意に置換されたフェニル基であり、
R^２は随意に置換された低級アルキル基、アラルキル基、チオアルキル基、アミドアルキル基、アミノ基、アミノアルキル基、ニトロ基、アジド基であり、Zと共に随意に置換された4-7員環の炭素環または複素環を形成し、あるいはR⁴と共に随意に置換されたフェニル基を形成し、
R³はR²と共に随意に置換されたフェニル基を形成するか、あるいは随意に置換されたフェニル基であり、
ZはH、低級アルキル基、置換アルキル基、チオアルキル基、二トリル基、アミノ基であるか、あるいはR²と共に随意に置換された4-7員環の炭素環または複素環を形成する。 Example 5 Further Examples of Compounds Although various compounds with immunomodulatory and / or anti-tumor activity have been disclosed herein, many of these compounds also have resonance modifying properties. These compounds can treat body surface lesions such as vagina and cancer, such as anogenital, genital, reproductive organs, or skin cancer. In some embodiments, the compound has the following general structure:

Ar in the formula includes an aryl group substituted with one or more electron withdrawing groups,
X is NH, CH ₂ , O, —, SO ₂ , or B,
Q is-(CH = CH) _m -CH-
Y and Y ′ are independently selected from N, CH, and B;
n is 0 or 1,
m is from 0 to 6,
R ¹ is H, a lower alkyl group, an optionally substituted phenyl group, or

R ⁴ in the formula is an optionally substituted phenyl group,
R ² is an optionally substituted lower alkyl group, an aralkyl group, a thioalkyl group, an amidoalkyl group, an amino group, an aminoalkyl group, a nitro group, an azide group, and a 4-7 membered ring optionally substituted with Z Forming a carbocyclic or heterocyclic ring or forming an optionally substituted phenyl group with R ⁴ ;
R ³ together with R ² forms an optionally substituted phenyl group or is an optionally substituted phenyl group,
Z is H, a lower alkyl group, a substituted alkyl group, a thioalkyl group, a nitrile group, an amino group, or forms an optionally substituted 4- to 7-membered carbocyclic or heterocyclic ring with R ² .

式中のR^５およびR^６は独立に、-OR⁷、-SR⁷、-SO₂N(R⁷)₂、-SO₂CN、-SO₂R⁸、-SO₂N₃、N₃、-CN、-N(R⁷)C(O)R⁸、-R⁷であり、
R⁷はH、低級アルキル基、糖、脂肪酸、アミノ酸、ペプチド、ヌクレオシド、または核酸であり、
R⁸は低級アルキル基またはC-グリコシドである。 Some embodiments include compounds of the structural formula above, where n is 1 or greater. In certain embodiments, such compounds are represented by the following structural formula:

R ⁵ and R ⁶ in the formula are independently -OR ⁷ , -SR ⁷ , -SO ₂ N (R ⁷ ) ₂ , -SO ₂ CN, -SO ₂ R ⁸ , -SO ₂ N ₃ , N ₃ , -CN, -N (R ⁷ ) C (O) R ⁸ , -R ⁷
R ⁷ is H, a lower alkyl group, a sugar, a fatty acid, an amino acid, a peptide, a nucleoside, or a nucleic acid,
R ⁸ is a lower alkyl group or a C-glycoside.

In other embodiments, the compound has the following structure:

式中のR^７およびR^８は独立に-OR⁵、-SR⁵、-SO₂N(R⁵)₂、-SO₂CN、-SO₂R⁶、-SO₂N₃、N₃、-CN、-N(R⁵)C(O)R⁶、-R⁶であり、
各R⁵は独立にHまたは低級アルキル基で、R⁶は低級アルキル基で、かつ、Ar、X、およびYは上記の通りである。 In certain embodiments, compounds having the general structure described above have n equal to 0. One embodiment of such a compound has the following structure:

R ⁷ and R ⁸ in the formula are independently -OR ⁵ , -SR ⁵ , -SO ₂ N (R ⁵ ) ₂ , -SO ₂ CN, -SO ₂ R ⁶ , -SO ₂ N ₃ , N ₃ ,- CN, -N (R ⁵ ) C (O) R ⁶ , -R ⁶
Each R ⁵ is independently H or a lower alkyl group, R ⁶ is a lower alkyl group, and Ar, X, and Y are as described above.

式中のR⁵はH、-OR⁶、-SR⁶、-SO₂N(R⁶)₂、-SO₂CN、-SO₂R⁷、-SO₂N₃、N₃、-CN、-N(R⁶)C(O)R⁷、または-R⁷であり、
R¹およびR⁶は独立にHまたは低級アルキル基で、
R²およびR⁷は独立に低級アルキル基で、かつArは上述のアリール基である。 Other compounds disclosed herein include those having the following structure:

R ⁵ in the formula is H, -OR ⁶ , -SR ⁶ , -SO ₂ N (R ⁶ ) ₂ , -SO ₂ CN, -SO ₂ R ⁷ , -SO ₂ N ₃ , N ₃ , -CN,- N (R ⁶ ) C (O) R ⁷ , or -R ⁷ ,
R ¹ and R ⁶ are independently H or a lower alkyl group,
R ² and R ⁷ are independently a lower alkyl group, and Ar is the aryl group described above.

式中のR¹は水素、ヒドロキシル基、2-ヒドロキシフェニル基もしくは4-ヒドロキシフェニル基、酢酸塩、ニトロソ基、リン酸塩、アジド基、二トリル基、アミノ基、ジメチルアミノ基、硫酸塩、メチルスルホン酸塩、リン酸塩、琥珀酸塩、または他の水素結合が可能な水溶性求電子基であり、R²はC₆H₄OH、C₆H₄N₃、C₆H₄CN、4-HO-C₆H₄-C₆H₄、C₆H₄OPO₂OH、C₆H₄OSO₂H、C₆H₄NH₂、C₆H₄NHMe₂、C₆H₄OSO₂Me、C₆H₄OCO(CH₂)_xCO₂H、またはC₆H₅Clであり、かつArは一つ以上のニトロ部分で置換されたアリール基をあらわす。或る実施形態では、ArはC₆H₃-2,4(NO₂)₂、C₆H₄-4(NO₂)、C₆H₄-3(NO₂)、またはC₆H₃-2,4(NO₂)₂を含む。或る特定の実施例では、R¹はOHで、R²はC₆H₄OH、かつXはC₆H₃-2,4(NO₂)₂である。 In certain embodiments, the compound is a mono or polyaryl mononitro- or dinitrophenyl hydrazone, such as

R ¹ in the formula is hydrogen, hydroxyl group, 2-hydroxyphenyl group or 4-hydroxyphenyl group, acetate, nitroso group, phosphate, azide group, nitryl group, amino group, dimethylamino group, sulfate, Methyl sulfonate, phosphate, oxalate, or other water-soluble electrophilic group capable of hydrogen bonding, R ² is C ₆ H ₄ OH, C ₆ H ₄ N ₃ , C ₆ H ₄ CN , 4-HO-C ₆ H ₄ -C ₆ H ₄ , C ₆ H ₄ OPO ₂ OH, C ₆ H ₄ OSO ₂ H, C ₆ H ₄ NH ₂ , C ₆ H ₄ NHMe ₂ , C ₆ H ₄ OSO ₂ Me, C ₆ H ₄ OCO (CH ₂ ) _x CO ₂ H, or C ₆ H ₅ Cl, and Ar represents an aryl group substituted with one or more nitro moieties. In some embodiments, Ar is C ₆ H ₃ -2,4 (NO ₂ ) ₂ , C ₆ H ₄ -4 (NO ₂ ), C ₆ H ₄ -3 (NO ₂ ), or C ₆ H _3- Including 2,4 (NO ₂ ) ₂ In one particular embodiment, R ¹ is OH, R ² is C ₆ H ₄ OH, and X is C ₆ H ₃ -2,4 (NO ₂ ) ₂ .

式中のR¹は水素、ヒドロキシル基、2-ヒドロキシフェニル基もしくは4-ヒドロキシフェニル基、酢酸塩、リン酸塩、アジド基、二トリル基、アミノ基、ジメチルアミノ基、硫酸塩、メチルスルホン酸塩、リン酸塩、琥珀酸塩、または他の水素結合可能な水溶性求電子基であり、R²はC₆H₅、C₆H₄OH、C₆H₅、C₆H₄N₃、C₆H₄CN、4-HO-C₆H₄-C₆H₄、C₆H₄OPO₂OH、C₆H₄OSO₂H、C₆H₄NH₂、C₆H₄NHMe₂、C₆H₄OSO₂Me、C₆H₄OCO(CH₂)_xCO₂H、またはC₆H₅Clであり、XはC₆H₃-2,4(NO₂)₂、C₆H₄-4(NO₂)、C₆H₄-3(NO₂)、またはC₆H₃-2,4(NO₂)₂であり、かつYは-O-、-S-、-CH₂-、-N-、―、-CHA-、または-CHOA-で、ここでAはアリール基、エステル基、アミド基、脂質、糖、またはペプチド残基である。特定の実施例では、R¹はOH、R²はC₆H₄OH、かつArはC₆H₃-2,4(NO₂)₂である。 In some embodiments, useful compounds are represented by the following structure:

R ¹ in the formula is hydrogen, hydroxyl group, 2-hydroxyphenyl group or 4-hydroxyphenyl group, acetate salt, phosphate salt, azide group, nitrile group, amino group, dimethylamino group, sulfate salt, methylsulfonic acid Salt, phosphate, oxalate, or other hydrogen-bondable water-soluble electrophilic group, R ² is C ₆ H ₅ , C ₆ H ₄ OH, C ₆ H ₅ , C ₆ H ₄ N ₃ , C ₆ H ₄ CN, 4-HO-C ₆ H ₄ -C ₆ H ₄ , C ₆ H ₄ OPO ₂ OH, C ₆ H ₄ OSO ₂ H, C ₆ H ₄ NH ₂ , C ₆ H ₄ NHMe ₂ , C ₆ H ₄ OSO ₂ Me, C ₆ H ₄ OCO (CH ₂ ) _x CO ₂ H, or C ₆ H ₅ Cl, where X is C ₆ H ₃ -2,4 (NO ₂ ) ₂ , C ₆ H ₄ -4 (NO ₂ ), C ₆ H ₄ -3 (NO ₂ ), or C ₆ H ₃ -2,4 (NO ₂ ) ₂ and Y is -O-, -S-, -CH _2- , -N-,-, -CHA-, or -CHOA-, where A is an aryl group, ester group, amide group, lipid, sugar, or peptide residue. In particular examples, R ¹ is OH, R ² is C ₆ H ₄ OH, and Ar is C ₆ H ₃ -2,4 (NO ₂ ) ₂ .

式中のR¹は水素、ヒドロキシル基、2-ヒドロキシフェニル基もしくは4-ヒドロキシフェニル基、酢酸塩、リン酸塩、アジド基、二トリル基、アミノ基、ジメチルアミノ基、硫酸塩、メチルスルホン酸塩、リン酸塩、琥珀酸塩、または他の水素結合可能な水溶性求電子基であり、R²はC₆H₄OH、C₆H₄N₃、C₆H₄CN、CH₃、4-HO-C₆H₄-C₆H₄、C₆H₄OPO₂OH、C₆H₄OSO₂H、C₆H₄NH₂、C₆H₄NHMe₂、C₆H₄OSO₂Me、C₆H₄OCO(CH₂)_xCO₂H、またはC₆H₅Clであり、XはC₆H₃-2,4(NO₂)₂、C₆H₄-4(NO₂)、C₆H₄-3(NO₂)、またはC₆H₃-2,4(NO₂)₂であり、YはH、(CH)_xCH₃（x = 0-12）、-S-CH₃、二トリル基、アミノ基、ニトロ基、アジド基、琥珀酸塩、またはアミド基であり、かつZはH、(CH)_xCH₃（x = 0-12）、-S-CH₃、二トリル基、アミノ基、ニトロ基、アジド基、琥珀酸塩、またはアミド基である。特定の実施例では、R¹はHまたはOH、R²はC₆H₄OHまたはC₆H₅、かつArはC₆H₃-2,4(NO₂)₂である。 In other embodiments, the compound has the following structure:

R ¹ in the formula is hydrogen, hydroxyl group, 2-hydroxyphenyl group or 4-hydroxyphenyl group, acetate salt, phosphate salt, azide group, nitrile group, amino group, dimethylamino group, sulfate salt, methylsulfonic acid A salt, phosphate, oxalate, or other hydrogen-bondable water-soluble electrophilic group, R ² is C ₆ H ₄ OH, C ₆ H ₄ N ₃ , C ₆ H ₄ CN, CH ₃ , 4-HO-C ₆ H ₄ -C ₆ H ₄ , C ₆ H ₄ OPO ₂ OH, C ₆ H ₄ OSO ₂ H, C ₆ H ₄ NH ₂ , C ₆ H ₄ NHMe ₂ , C ₆ H ₄ OSO ₂ Me, C ₆ H ₄ OCO (CH ₂ ) _x CO ₂ H, or C ₆ H ₅ Cl, where X is C ₆ H ₃ -2,4 (NO ₂ ) ₂ , C ₆ H ₄ -4 (NO ₂ ), C ₆ H ₄ -3 (NO ₂ ), or C ₆ H ₃ -2,4 (NO ₂ ) ₂ , Y is H, (CH) _x CH ₃ (x = 0-12), -S -CH ₃ , a nitrile group, an amino group, a nitro group, an azide group, an oxalate group, or an amide group, and Z is H, (CH) _x CH ₃ (x = 0-12), -S-CH ₃ , nitrile group, amino group, nitro group, azide group, oxalate Or an amide group. In particular examples, R ¹ is H or OH, R ² is C ₆ H ₄ OH or C ₆ H ₅ , and Ar is C ₆ H ₃ -2,4 (NO ₂ ) ₂ .

式中のGは-CH₂-、-C(O)-、-N-、―、-O-、または-S-をあらわし、nは0または1で、R⁷およびR⁸は独立に、-OR⁵、-SR⁵、-SO₂N(R⁵)₂、-SO₂CN、-SO₂R⁶、-SO₂N₃、N₃、-CN、-N(R⁵)C(O)R⁶、-R⁶であり、Arは一つ以上の電子求引基で置換されたアリール基を含み、
XはNH、CH₂、O、SO₂、またはBで、
Qは-(CH=CH)_m-CH-で、
YおよびY'は独立にN、CH、およびBから選択され、
各R⁵は独立にHまたは低級アルキル基で、かつR⁶は低級アルキル基である。 Further useful compounds are disclosed in WO 94/05276 ('276 publication), which is incorporated herein by reference. The compounds disclosed in the '276 publication are currently known to have immunomodulatory and / or resonance modifying activity. An embodiment of such a compound has the following structure:

G in the formula represents —CH ₂ —, —C (O) —, —N—, —, —O—, or —S—, n is 0 or 1, R ⁷ and R ⁸ are independently -OR ⁵ , -SR ⁵ , -SO ₂ N (R ⁵ ) ₂ , -SO ₂ CN, -SO ₂ R ⁶ , -SO ₂ N ₃ , N ₃ , -CN, -N (R ⁵ ) C (O ) R ⁶ , -R ⁶ , Ar comprises an aryl group substituted with one or more electron withdrawing groups,
X is _{NH, CH 2, O, SO} 2 , or B,,
Q is-(CH = CH) _m -CH-
Y and Y ′ are independently selected from N, CH, and B;
Each R ⁵ is independently H or a lower alkyl group, and R ⁶ is a lower alkyl group.

式中のR²はH、低級アルキル基、もしくは随意に置換されたフェニル基であり、R³はHであるか、またはR²と共に、随意にアリール基と縮合する、随意に置換された複素環または炭素環を形成し、R⁴、R⁵およびR⁶は独立にH、-OH、および-OR⁷から選択される。 Certain embodiments of the compounds disclosed herein are represented by the following structural formula:

R ² in the formula is H, a lower alkyl group, or an optionally substituted phenyl group, and R ³ is H or an optionally substituted heterocycle that optionally condenses with R ² together with an aryl group. Forming a ring or carbocycle, R ⁴ , R ⁵ and R ⁶ are independently selected from H, —OH, and —OR ⁷ ;

Many structures and synthetic methods of the described compounds have already been described in US Pat. No. 4,732,904 and are incorporated herein by reference. Some compounds not mentioned in that patent include 10-[(2,4-dinitrophenyl) hydrazono] -1,2-dihydroxy-10H-anthracen-9-one (DNPA). The synthesis of this compound is carried out by dissolving 0.1 mol of 1,2-dihydroxy-9,10-anthraquinone in 50 ml ethanol and 5 ml sulfuric acid. The solution was heated to 60 ° C. with gentle stirring and 50 ml ethanol containing 0.1 mol of 2,4-dinitrophenylhydrazone was added slowly with continued stirring. The solution was heated for 2 hours and then cooled in the refrigerator. DNPA crimson crystals appear, the melting point is 240-242 ° C., and the yield is 92%. When the product was analyzed as C ₂₀ H ₁₂ N ₄ O ₇ , C was 63.98, H was 3.46, N was 11.13 and the NMR spectrum and mass spectrum were consistent. Additional compounds suitable for use in the compositions and methods of the present disclosure are disclosed in WO 2004/078174, issued 16 September 2004. The compounds disclosed in this publication are incorporated herein by reference.

  Further examples of resonance modifiers are provided in Example 13.

Example 6 Formulations for Administration to a Subject Drugs are provided in many forms, such as crystals, pellets, or gels / creams. The drug can be implanted subcutaneously or applied to the epithelial surface of a patient who appears to have a compromised immune system or who has come into contact with a lethal or toxic substance capable of down-regulating the immune system can do. The drug can be incorporated into any shape (such as a patch or a two-dimensional or three-dimensional substrate) that is in electromagnetic contact with the target site. “Electromagnetic contact” means close enough to exert an immunomodulatory effect as described herein. In some examples, the agent is applied to the lesion being treated (such as a tumor or dysplastic epithelium) or applied over the lesion. If desired, the drug can be chemically modified, for example, to form a polymer. Agents are also useful for discovering cancer-related changes in immune function, such as when there is a deficiency in the autoregulation of the immune system.

  In one example, A-007 pellets were prepared by compressing 50 mg of pure compound to a 16 gauge size. Bulk A-007 was prepared using the GLP / GMP procedure without the addition of additives. Depending on the solubility characteristics of the 100% A-007 pellets, additives (stearic acid, povidone, etc.) or other pharmaceutically acceptable carriers may be added. Pellets can be produced at doses of, for example, 25 mg, 50 mg, and 75 mg. Adjustment of the pellet concentration can be made depending on the observed physical properties (such as solubility characteristics) and animal toxicity. The therapeutically effective amount can be determined by known means, and the dosage may vary depending on the condition being treated or the severity of the disease.

The drug can also be provided in the form of a gel, such as a propylene glycol / methyl cellulose formulation, for application to a target site (such as a body part) where it is desired to concentrate the immune response. The gel can be provided in a tube, such as 50 mg of active agent suspended within the gel. These gels are particularly useful for application to skin or other epithelial surfaces (eg, chest wall skin, back, foot skin, face, or anogenital region, or cervical mucosa). In some embodiments, the gel is dispensed from a long application tube (such as a rectal or vaginal applicator) and applied, for example, rectally or transvaginally.

  Alternatively, the pellet can be inserted into a target site such as the chest wall. In other examples, the drug is suspended in tetrahydrofuran (THF) or other non-toxic aerosol and introduced into the tracheobronchial or oropharyngeal site to treat oral or upper respiratory tract cancers.

  The pellets have been found to promote the deposition of uric acid crystals, which can cause painful nodules. Thus, although application to the skin surface is preferred over insertion into the skin, both approaches cause the immunomodulation described herein.

Example 7 External Monitoring Device for Immune Activity As described above, the described agents can both monitor immune properties and / or up-regulate in the presence of sudden behavioral modifications related to pathology .

  In its simplest form, the monitoring device is a device that detects changes in the electromagnetic properties of the drug, such as a resonance modifier that interacts with a biological system. The interaction can be observed either in vitro or in vivo. Accordingly, the monitoring device can be applied to a subject or a biological sample (such as tissue culture) taken from the subject for further analysis.

  Resonance modifier (A-007) is present on the skin of lymphoma patients as 0.25% propylene glycol / methyl cellulose gel (0.5 g). Although shown as a gel, this can be applied on the epithelial surface or skin (or other surface), or inserted into it with a 16 gauge trocar, or cream or single crystal (50 mg) or It can also be applied as a pellet (50-100 mg). In one embodiment, the drug is applied to the skin as an orange gel, and a 2x3 Tegaderm® patch with two electrode tabs of the type used as electrocardiogram leads being adhered to the skin surface at a distance. Covered with. The electrodes are placed on both sides of the gel in such an arrangement that the potential difference between the gels can be measured. Then two probes of the voltmeter are connected and one probe is connected to each electrode tab. The voltmeter is preferably coupled to an oscilloscope that measures voltage changes over time so that the amplitude of the detected voltage can be viewed over time. In a particular embodiment, the voltmeter is a Fluke® Scopemeter 192/196/199, which can filter the voltage waveform to remove background voltage, and further observe and record on a computer. Have software that can measure and analyze. Thus, changes in voltage amplitude over time are observed to monitor the immune response to the resonance modifier.

  After setting the reference amplitude and / or frequency when the resonance modifier is applied to the skin, the electrodes are subsequently connected to an oscilloscope to monitor the amplitude and frequency of power transmission from the test site. For resonance modifiers such as A-007, the amplitude of the voltage change over time increases.

  After placing the patch on a normal subject for 7 days, the tip of the voltmeter probe was again connected to the electrode lead and the voltage change over time was recorded. The electrical activity associated with aggregation and increased mitotic activity caused by the migration of activated dendritic cells to the target site of the patch was measured. The amplitude of the waveform (measured from zero) increased by approximately 25% from 3 to 4 volts.

  Although this example showed the measurement of electrical activity changes using a voltmeter, the electrodes can also be detected with an external laser beam detector that detects changes in local chemical-cell interactions. . Since chemicals can conduct electrical energy via resonance, chemicals provide signals for aspects of cell-cell interaction. For example, a molecular scanner such as a laser can be used for activating a chemical substance or for analysis at its absorption wavelength (404 nm for A-007). One such molecular scanner is the Edmund Industrial Optics solid-state tunable laser (wavelength 425 nm), which is programmed to record or create intrinsic excitations related to A-007 resonances and cell-cell interactions. ing. These high performance lasers may be programmed to recognize nano-scale A-007 spectral characteristics, much like bar code scanners.

Example 8 Enhancement of Immune Function Modulation The disclosed compounds also provide a relatively simple way to enhance immune function, for example, in a subject suffering from immune deficiency. Such subjects may be suffering from, for example, immunodeficiency (such as HIV / AIDS) or lack of toxin-induced immunity (such as leukopenia induced by anti-tumor chemotherapeutic drugs or environmental toxins). Other subjects have the type of immunodeficiency often found in patients with malignant tumors. Alternatively, subjects may have objective laboratory values for immune deficiency, regardless of whether the primary cause of immune deficiency is known. The compound can serve as a coupling agent or contact with the immune system and can not only monitor the immune system, but can also be therapeutically controlled.

  As already indicated, by introducing the drug into or near the target site, it is possible to direct the target immune response to the target site (such as near the tumor). Exposing the resonance modifier to an electromagnetic field that stimulates the electromagnetic properties of the agent stimulates the resonance modifier. For example, the drug can be exposed to an external magnetic or electric field. An example of such an electromagnetic field provides the highest concentration of the resonance form of the resonance modifier (such as one of the forms of A-007 shown in FIG. 3) in order to provide maximum contact with the cell receptor surface. A signal having such a frequency is sent or adjusted to that frequency. In this example, the laser is selected to apply an electromagnetic field in the form of laser energy having the same wavelength (400-450) as indicated by A-007. A-007 does not have laser oscillation characteristics, but a programmed laser can recognize the spectral characteristics of A-007. A simple magnetic field created by a 2 AA or 3 V battery can activate the surface gel of A-007 nonspecifically, or in the case of 9 V it can activate a subcutaneous pellet. More specialized tunable lasers can scan the region and are more specific for the optimal electromagnetic field to apply. By choosing a preferred frequency, the amplitude of the voltage change over time seen with the resonance modifier increases, which in turn enhances or activates the immune response.

  To treat tumors that are present in both the dermis and epidermis, a conjugated aromatic compound is applied to the skin surface above the target site containing the tumor. While the compounds attract dendritic cells and lymphocytes to the target site, tumors can often circumvent normal immune surveillance and are useful for further stimulating the immune response through high activation of resonance modifiers. A magnetic probe placed over the site where the compound is applied to the skin provides such high activation. The magnetic probe is connected to a device that includes an oscillating circuit, a frequency counter, a voltage source, and an oscilloscope. The device is activated to apply a magnetic pulse to the compound to enhance its resonance modifying properties.

  An example of a suitable magnetic pulse generator is The Magnetic Pulser, which is a High Intensity Momentary Time-Variant Pulsed DC Magnetic Field Therapy Generator MODEL #: MPG5 available from Health Canada. The Magnetic Pulser (MPG5) is designed to generate a strong (~ 43,133 Gauss), instantaneous (~ 2.5mS), pulsed DC magnetic field and can be used to stimulate resonance modifying compounds. The magnetic pulse can be applied for a duration sufficient to enhance the activity of the compound. For example, a pulsed DC magnetic field can be applied over 5-60 minutes.

  A simple magnetic field created by a 2 AA or 3 V battery activates the surface gel of A-007, or a 9 V battery activates a subcutaneous pellet. More specialized tunable lasers can scan a region (such as a region where a compound is locally applied) and can provide a more specific indication of the applied electromagnetic field. By choosing a preferred frequency to apply the field, the amplitude of energy created by the resonance modifier is maximized. The duration for which the electromagnetic field is applied depends on the subject's immune status, and a longer stimulation period is useful for subjects with a poorer immune status. Although a common magnetic field is used, the preferred approach for a wide range of treatments is to use a wavelength or energy specific tunable transmitter. The effect of treatment is assessed by monitoring immune status with readily available techniques such as peripheral T-cell flow cytometric analysis. These readily available and convenient tests provide further guidance in selecting the applied electromagnetic field characteristics for a particular resonance modifier.

  In one embodiment, the applied electromagnetic stimulus is an electric field of the type utilized in US Pat. No. 6,190,893, which is incorporated by reference. The apparatus for electrical stimulation includes a gold wire cathode that extends along one end of the target site and a silver wire anode that is disposed along the opposite end of the target site. EG & G Princeton Applied Research Potentiostat / Galvostat Model 263A (Oakridge, TN) is used as a constant potential source. Electrical stimulation can be applied at a steady potential of 100 mV for a period of more than one hour.

  Many other devices are available for applying an electromagnetic field to the resonance modifier in human body or tissue culture. For example, PCT Publication WO 02/102457, which is incorporated herein by reference, is a device that includes a transducer and a generator that applies an AC signal to the transducer, with a fundamental electromagnetic field between 0.1 Hz and 4000 Hz. An apparatus is disclosed that is made at frequency. A similar device is also shown in US Pat. No. 5,968,527, which is incorporated by reference.

  In some situations, it is preferable to use a bioimpedance device to both provide stimulation to the resonance modifier and to monitor the electrical properties of the target site to which the resonance modifier has been applied. A direct bioimpedance measurement system uses a constant current source to generate a continuous, constant amplitude and constant frequency current through a human or animal body segment for the purpose of measuring tissue conductivity. A frequency in the range of 30 KHz-30 MHz is usually used. The impedance to the continuous current flow in the body segment creates a potential difference across the body segment and the bioimpedance meter measures the impedance in the body segment. Examples of bioimpedance devices are shown in WO 01/76475, US Patent No. 4,805,621, and US Patent No. 5,529,072. The bioimpedance probe is placed over the target site to which the resonance modifier is applied, and a potential difference is introduced across the target site. The change in bioimpedance is also used for monitoring the state of the subject's immune system. Increased bioimpedance is considered to be an improvement in immune status.

Example 9 Use of Disclosed Compounds in the Treatment of Lymphoma As described in Example 7, A-007 0.25% gel (0.5 g) was applied to the skin of a 42 year old lymphoma patient, and the patient's peripheral lymph There was an improvement in sphere shape (measured by total lymphocyte count) and an increase in the amplitude of the sigma wave, suggesting increased skin activity associated with A-007. Detection was performed using a Fluke® ScopeMeter oscilloscope and a PC program (192/196/199) that can be recorded and that can remove background noise from the environment. In this particular example, a 25% increase in voltage amplitude was considered a positive modification of immunity, consistent with improved lymphocyte shape.

[Example 10] Use of compounds in the treatment of cervical cancer A-007 0.25% gel (2 g) was applied daily for 5 days to a 23-year-old woman suffering from early cervical carcinoma in situ. The dramatic healing that continued was seen. Epidermal cancer cells disappeared, and normal appearance of epithelial tissue was obtained with the influx of CD45 + T-lymphocytes. The response is associated with an increase in T-lymphocyte type tissue and the disappearance of cancer cells.

Example 11 Protein Tyrosine Phosphatases Human protein tyrosine phosphatases (PTPs) are a large variety of protein families present in all eukaryotes. Each PTP consists of one or more conserved domains characterized by an 11 residue sequence motif containing cysteine and arginine residues, the latter being known to be essential for catalytic activity. The sequence of PTP does not share similarities with serine or threonine, acid or alkaline phosphatase. The structural diversity in the PTP family is mainly derived from the type of non-catalytic sequence attached to the NH ₂ -or COOH-terminus of the catalytic domain. There are many PTPs involved in intracellular phosphorus metabolism and domains. The diversity of the extracellular region probably reflects the type of ligand that PTPs are exposed to catalyze phosphate transfer.

  Extracellular PTPs are one type of PTP receptor for surface recognition and adhesion molecules for leukocyte cell surface recognition. PTPs are not only associated with human cells, but are also present in prokaryotes and viruses, and bacteria. In the pathogenic bacterium Yersinia, the pathogenic factor of intestinal plague, Yop2b tyrosine-specific PTP, is an essential toxicity determinant.

  Numerous studies have shown the importance of PTPs in physiological processes. Phenotypic abnormalities and abnormal proliferative behavior of T- and B-lymphocytes, granulocytes and macrophages are considered important issues in the progression of cancer and autoimmune diseases.

  The catalytic domain of PTPs has been described in crystallographic studies and is reviewed in Z. Jia, et al., Structural Basis for Phosphotyrosine Peptide Recognition by Protein Tyrosine Phosphatase 1B, Science 268: 1754-1758, 1995. This reference and Z. Xu et al., Negative regulation of CD45 by differential homodimerization of the alternatively splied isoforms, Nature Immunology 3: 764-771, 2002 disclose various types of cells, bacteria and viruses that express PTPs. ing. This example outlines the interactions and effects of aryl hydrazones as PTP modifiers.

  An overview of 4,4'-dihydroxybenzophenone-2,4-dinitrophenylhydrazone (A-007) and its structural properties (Figures 1 and 3) show that it does not satisfy Lipinsky's "rule of five" Yes. “Rule of Five” means that for absorption or penetration, the drug has a hydrogen bond donor of 5 or less, a hydrogen bond acceptor of 10 or less, a molecular weight of 500 or less, and / or a calculated log P of 5 or less. (CAChe Group, Fujitsu, Beaverton, OR). Clearly, as can be seen by crystallography, A-007 and other hydrazones have more than 5 hydrogen bond donors (Klein, CL, Gray. D., and Stevens, ED, Crystal and molecular structures of benzophenone phenylhydrazone derivatives with anticancer activity, Structural Chemistry 4: 377-383, 1993). Thus, since the drug is not substantially absorbed through the applied skin, the external binding to lymphocytes and cancer cells described in this patent is a structure-behavioral observation of the A-007 arylhydrazone. Matches. Therefore, as expected from the electromagnetic effect, the effect is obtained at a distance.

As shown in FIG. 1, A-007 fits well with the CD45 + PTP subtype extracellular catalytic receptor. In addition, other compounds disclosed herein, including the following structures, are well suited to these receptors.

Cervical cancer (transformed through HPV-induced mutation) cell death in the presence of A-007 crystals. This supports A-007's ability to catalyze HS-SH dimerization of cysteine residues through resonance to enhance receptor and cell death. This was different for patients with cervical cancer (Table 2). 0.25% A-007 topical gel was applied intravaginally at a dose of 2 grams daily for 5 days. A-007-induced cell death was not observed in benign fibroblasts or normal epithelial cells that did not contain HPV or were not deformed by HPV. A compound containing a resonance modifier, such as A-007, is applied topically to the cervical / supavaginal membrane of HPV-infected patients to remove cancer cells, further remove viral particles, and CD45RO + CD4 + and CD8 + T -Upregulation of lymphocytes was observed. The greatest therapeutic effect of topical application of A-007 is induction of cell death in HIV-infected cells (Table 2 and Figures 10 and 13). As shown in Table 2, A-007 increases the presence of T-lymphocytes, specifically CD45 +, CD123 +, CD4 + and CD8 + cells. In particular examples, an increase in CD45 +, CD4 +, and CD8 + cells was particularly noted. In particular, an increase in CD45RO +, CD45RA + and CD45RB + was noted.

  Although shown as a gel in this example, the disclosed compounds can be applied to the epithelial surface or skin (or other surface) for short-term treatment, or they can be used with a 16 gauge trocar. It can also be applied as a cream, single crystal (50 mg) or pellet (50-100 mg) that can be inserted inside.

  HeLa cancer cells (HPV +), when cultured with A-007 crystals for 24 hours, lead to necrosis and cell death. Transparent normal cancer cells migrate to the crystal. Then, through the cell membrane chemistry interface, cells down-regulate PTP receptor regulation, darkening with concentration changes associated with DNA aggregation, leading to death (down-aggregated cells). The attractive force a crystal has on a cell is related to intramolecular resonance and its associated magnetic form.

  Naive T-lymphocytes are also activated in the presence of A-007 along with activated lymphocyte + aggregation.

  Thus, A-007 and other disclosed compounds, such as the hydrazone described, can not only play a unique role in sensitizing HPV positive epithelial cells to attack by activated T-cells ( Table 2), directly affects HPV positive cells and A-007, resulting in cell and virus death (Table 2). In the latter table, the patient is no longer infected with the virus after treatment, indicating that the drug also has antiviral properties. Thus, A-007 not only upregulates CD45 + activity of T-lymphocytes, but also both deregulates the outer membrane and removes HIV-infected cells.

  These findings indicate that both HPV-infected cervical epithelial cells and T-lymphocytes contain extracellular membrane-bound PTPs with which the disclosed agents, including A-007, can interact. HPV viruses contain active extracellular PTP receptors, which can be deregulated and cell death when exposed to the compounds disclosed herein. Regardless of the actual mechanism, the disclosed compounds have been found to have effective antiviral activity against HPV. In addition, T-lymphocytes are activated by resonance modifiers such as A-007 and are thought to promote the presence of surface membrane CD45 + RO PTPs, which occurs through HS-SH formation in the catalytic receptor site it is conceivable that. CD45 + RA T-lymphocytes represent quiescent or naïve cells that cannot attack foreign cells, so conversion to the CD45 + RO marker represents activation of the cytotoxic arm of the cellular immune system . As shown in Table 2, the resonance modifier has substantially antiviral (anti-HPV) activity and antitumor activity. The proportion of CD45 + RO cells increases substantially after drug treatment.

  Cells expressing PTPs, such as plague pathogens, or PTP + cells infected with pathogens (such as HPV-infected epithelial cells) can also aggregate with A-007. This aggregation is thought to occur by cysteine oxidation and disulfide bond formation. This bond formation prevents cell integrity and cell death-virus release. In the presence of A-007, viral PTP cell membrane autooxidation occurs with virus inactivation. Most patients in Table 1 also have HPV + proliferating cervical cells with HPV-induced nuclear changes (CIN) that disappear with A-007 treatment. This further emphasizes that the disclosed compounds, including resonance modifiers such as A-007, have the ability to destroy not only malignant cells, but also benign and potentiated cells infected with HPV.

  Anticancer and antiviral activity in hydrazone analogs, such as 2,6-dibenzylidenecyclohexanone-2,4-dinitrophenylhydrazone, improved the binding affinity and upregulation of PTP CD45 receptor, but exposure to ultraviolet light Also promoted through. Ultraviolet light induces a Diels-Alder reaction between A-007 and the diimine site of arginine, as well as adduct formation with the cysteine HS- site, leading to upregulation of the CD45 receptor (FIG. 1). Thus, the immunomodulating activity, antiviral activity, and antitumor activity of such resonance modifiers can be enhanced by exposing the compound to ultraviolet light.

[Example 12] Antitumor and immunomodulating properties of 4,4'-dihydroxybenzophenone-2,4-dinitrophenylhydrazone (A-007)
4,4′-Dihydroxybenzophenone-2,4-dinitrophenylhydrazone (A-007, compound 1 in FIG. 12) produced significant anticancer activity in patients with anal squamous cell carcinoma (Table 3). A topical gel containing 0.25% A-007 (2 grams of gel per day) was applied intraanally daily for 5 days. The observed results are shown in Table 3.

Example 13 Further examples of disclosed compounds
2,6-Dibenzylidenecyclohexanone-2,4-dinitrophenylhydrazone (BDP-DNP) is a cell growth of malignant squamous cell carcinoma (SCCA-HM) that grows in culture (pre-post Rx) As shown by the effect of BDP-DNP on ATP, it has 100 times greater immunomodulating anticancer activity than A-007. After adding BDP-DNP (0.4 mcg / mL) in tissue culture medium (LR Morgan, US Pat. No. 5,270,172) and culturing for 24 hours, lymphocytes received a remarkable stimulus (a large white population of activated T-lymphocytes) And further cancer cell destruction happened. No cancer cells were found.

Example 14 Characteristics of some active compounds As disclosed in the previous examples, the compounds disclosed herein provide both diagnostic and therapeutic contacts with the immune system, and Have antitumor and antiviral activity (especially against HPV and HPV infected cells, such as vaginal and cervical cancers associated with HPV infection).

  The compounds are disclosed to have one or more of the following characteristics, for example, having at least three, five, or all of the following characteristics.

  Specific attraction of CD45 + lymphocytes (eg CD45RO + and CD45RA + lymphocytes) to resonance-modifying compounds, either in culture or when applied to the surface of a living organism (such as skin).

  For example, up-regulation of CD45RA + to CD45RO + lymphocytes when applied to the epithelium of a subject.

  Ownership of intrinsic electromagnetic properties, for example, the ability to form a single wire of agglomerated crystals between positive and negative charge points when the crystals are deposited, or release of electromagnetic energy.

  Ownership of properties that affect lymphocyte migration, eg, induction of aggregation of naive lymphocytes in culture.

  Certain embodiments of the compound are also characterized by the ability to increase the amplitude of the alternating voltage wave measured from the compound after being applied to the skin of the individual. In certain embodiments, the increased amplitude is reduced in immunocompromised subjects, such as lymphocyte dysfunction.

  In certain embodiments, substantially no skin absorption is observed when the compound is applied to the skin. In some embodiments, such compounds do not satisfy Lipinsky's “rule of five”. Such compounds remain substantially entirely on the skin surface when applied to the skin without transdermal flow.

  Potential resonance modifiers can also be selected based on the chemical structure presenting the resonance modification, ie, the structural properties described earlier in this specification. For example, polyaryl compounds having electronegativity such as Casodex (registered trademark), Naprelan (registered trademark), Eulexin (registered trademark), and Bextra (registered trademark) can be selected.

Once candidates are selected as potential resonance modifiers, they can be easily analyzed to determine whether they induce lymphocyte blastogenesis and activate lymphocytes. Such an analysis is found in Morgan et al., Anticancer Research 13: 1763-1768 (1993), which is incorporated by reference. See also Janossy et al., Clin. Exp. Immunol. 14: 581-596 (1973). Briefly, [ ³ H] thymidine incorporation was measured in cultured lymphocytes. Lymphocytes were cultured in RPMI-1640 tissue culture medium supplemented with 10% fetal bovine serum (FBS). Following stimulation with the resonance modifier, tritiated thymidine (2-20 μCi / ml, specific activity 2 Ci / mmol) was added to the cells and cultured together. [ ³ H] thymidine incorporation was stopped after the selected time by adding cold 10% trichloroacetic acid and the radioactivity of the specimen was measured with a scintillation counter. % Lymphoblasts were determined by counting the number of blasts per high powered field (hpf), while% activation was determined by tritiated thymidine incorporation. The voltage increase was measured with an oscilloscope as in Example 7.

  This procedure represents a correlation between increasing the amplitude of the voltage signal and promoting activation of lymphocytes. In particular, a greater amplitude increase was observed with a more active increase in lymphocyte activation. This analysis can be used to quickly select candidate resonance modulators for immune activation. For example, using this analysis, an increase in voltage amplitude of 20% or more (eg, an increase in amplitude of 50% or more) can be used as a basis for resonance modifying activity in the immune system. Resonant modification candidates that meet such criteria (eg, greater than 20% amplitude increase) are then selected for further use and research.

Example 15 Topical Formulation The compounds disclosed herein can be formulated as a topical formulation for application to the skin as an immunostimulant, antiviral agent, or antitumor formulation.

  In certain examples, the compound comprises 2,6-dibenzylidenecyclohexanone-2,4-dinitrophenylhydrazone (BDP-DNP). This is a topical formulation for application to epithelial surfaces, for example by application to malignant epithelia such as urogenital tumors, such as anus, vagina, or cervical neoplasms (eg cervical CIN). Be put in.

  The compounds can be utilized in methods of treating tumors (neoplasms) by administering a therapeutically effective amount of an agent to an affected patient to induce regression or elimination of tumor cells. In certain examples, administration of BDP-DNP includes topical administration of BDP-DNP to the tumor, although other forms of administration such as oral, inhalation, infusion, or subcutaneous administration may also be included.

[Example 16] Formulation for inhalation
A compound (A-007 in this example) in a 5% solution of tetrahydrofuran (THF) is released into the mouse's respiratory system by releasing the aerosol into the closed room of the mouse for 30 seconds under a pressure of 15 psi. Sprayed. As a result of the treatment, none of the treated mice died. Five days later, the mice were sacrificed and the lung system was examined by excising the trachea and lungs. Histological examination of the specimen revealed increased bronchial epithelial lymphocyte infiltration (CD8 + and CD4 +). The same pattern of T-cell modification was seen with topical application to the neck and skin. The control group was treated with THF alone and no increase in lymphocyte infiltration was observed.

Example 17 Active Agent-Receptor Interaction This example describes a modeling test used to assess the binding of compounds disclosed herein to receptors such as CD45. These tests were performed using Sybyl 6.9.2 (Tripos Inc, St Louis, MO). Compound models were made from Sybyl fragments and then optimized using the MMFF94 force field (with MMFF94 charge). These models were then docked to the 1BZC crystal structure active site using FlexX 1.13 and the Drug Score scoring function. FlexX performs incremental fragment based docking, where various base fragments of the ligand are docked, and then the rest of the ligand is assembled from this basic fragment arrangement . Thus, all possible ligand orientations and conformations can be sampled.

これらの試験において、示された化合物はA-007よりも良い結合スコアを持ち、この主な理由は、上記の化合物のジヒドロキシビフェニル部分と密接に接触するポケットへの入り口でARG47によってより強く保持されるためである。 FIG. 19 shows the docking of the following compounds with CD45 according to this method.

In these tests, the indicated compound has a better binding score than A-007, which is mainly retained by ARG47 at the entrance to the pocket in close contact with the dihydroxybiphenyl moiety of the above compound. Because.

  The illustrated compounds are also found to aggregate where they bind to CD45 and exert direct cytotoxic activity.

Example 18 General Procedure for the Preparation of Hydrazone Method 1: Substituted phenylhydrazine (0.148 mol) was suspended in MeOH (300 mL) at 50 ° C. and concentrated sulfuric acid (20 mL) was stirred at 50 ° C. added. After the hydrazine was dissolved, MeOH (300 mL) containing a substituted benzophenone or aryl ketone (0.1 mol) was added with stirring at 50 ° C. for 10 minutes or more, and then further stirred for 30 minutes. The solvent was evaporated (75%), water (500 mL) was added and the precipitate was washed with 3% aqueous NaHCO ₃ . Recrystallization was performed with EtOH, MeOH, or acetic acid. Complete elemental and spectral analyzes were performed on all materials.

  Method 2: The second general method is performed as follows. The substituted aryl hydrazine (2.7 mmol) was dissolved in 15 mL MeOH and 2.1 mmol ketone was added with stirring. Dowex 50W-50X2-100 cation exchange resin (0.5 g) [Dow Corp., Baton Rouge, LA] was added. The suspension was refluxed at 75 ° C. for 1 hour on a water bath. The mixture was filtered to hot under water vacuum pressure and the resin was washed 3 times with 0.5 mL MeOH. The contents of the collection flask were cooled to room temperature and 20 mL of distilled water was added. The colored precipitate was filtered, dried and recrystallized from EtOH or MeOH. Typical yield was in the range of 80%.

N-benzhydrylidene-N ′-(2,4-dinitrophenyl) hydrazine was prepared using Method 1 and yielded yellow crystals with a recovery of 82% from EtOH or MeOH. mp is 232-234 ° C, IR is 3295 (NH), 2928 (Ar-H), 1612 (C = N), 1590, 1504, 1416, 1334, 1127, and 1091, 834, 777, 698, and 608 cm ^-1 and ¹ H-NMR (DMSO, 400 MHz) are δ11.25 (1H, s, NH), 9.07 (1H, s), 8.37 (1H, d, J = 8 Hz), 8.22 (1H, d, J = 8 Hz), 7.67 (4H, m, J = 8 Hz), 7.41 (6H, m, J = 8 Hz), C ₁₉ H ₁₄ N ₄ O ₄ elemental analysis calculated C, 62.98, H 3.86, N, 15.46, the measured values were C, 63.17, H, 4.06, N, 15.49.

4-Hydroxybenzophenone-2,4-dinitrophenylhydrazone was prepared using Method 2 and was obtained as red needle crystals that were recrystallized from EtOH in 80% yield. mp is 224-225 ° C, IR is 3541 (NH), 3271 (NH), 3098 (OH), 2905 (Ar-H), 1621 (C = N), 1591, 1511, 1418, 1329, 1276, 1130, 1083, 829 and 698 cm ⁻¹ , ¹ H-NMR (DMSO, 400 MHz) is δ11.23 (1 / 2H, s, NH), 10.99 (1 / 2H, s, NH), 10.1 (1H, s, "broad" OH), 8.8 (1H, s), 8.41 (1H, d, J = 8 Hz), 8.19 (1H, d, J = 8 Hz), 7.67 (2H, m, J = 8 Hz), 7.48 (3H, m, J = 8 Hz), 7.28 (1H, dd, J = 8 Hz), 7.04 (1H, dd, J = 8 Hz), 6.84 (1H, dd, J = 8 Hz), C ₁₉ H The calculated values of elemental analysis of ₁₄ N ₄ O ₅ were C, 59.07, H, 3.62, and the actually measured values were C, 59.71, H, 3.84.

2,2 ′, 4,4′-Tetrahydroxybenzophenone-2,4-dinitrophenylhydrazone was synthesized as a crimson crystal recrystallized from EtOH with a yield of 75%. mp is 290-291 ° C, IR is 3631 (NH), 3491 (NH), 3272 (OH), 2905 (Ar-H), 1614 (C = N), 1518, 1415, 1322, 1206, 1143, 1107 , 974, 864 and 827 cm ⁻¹ , ¹ H-NMR (DMSO, 400 MHz) is δ11.59 (1H, s, OH), 11.15 (1H, s, NH), 10.12 (2H, s, OH), 9.91 (1H, s, OH), 8.85 (1H, s), 8.48 (1H, d, J = 8 Hz), 8.19 (1H, d, J = 8 Hz), 7.61 (1H, d, J = 8 Hz) ), 6.95 (1H, dd, J = 8 Hz), 6.8 (1H, dd, J = 8 Hz), 6.57 (1H, s), 6.46 (1H, dd, J = 8 Hz), 6.38 (1H, s ), 6.31 (1H, dd, J = 8 Hz), calculated values for elemental analysis of C ₁₉ H ₁₄ N ₄ O ₈ are C, 53.52, H, 3.28, N, 13.14, measured values are C, 51.79, H 3.19, N, 13.36.

1,2-Dihydroxy-10-anthraquinone-2,4-dinitrophenylhydrazone was synthesized from base quinone and 2,4-DNP using General Method 1. The yield was 92%. The reaction mixture was refluxed for 48 hours. The compound precipitated upon cooling of the reaction and was recrystallized as red microcrystals from dioxane / hexane. mp is 240-242 ° C, IR is 3502 (NH), 3287 (NH), 3113 (OH), 2925 (Ar-H), 2559 (Ar-H), 1618 (C = N), 1595, 1501, 1441 , 1335, 1298, 1135, 1091 and 781 cm ^-1 , ¹ H-NMR (DMSO, 300 MHz) is δ11.97 (1H, s, NH), 8.86 (1H, s), 8.47 (1H, d, J = 6), 8.38 (1H, s), 8.24 (1H, d, J = 12 Hz), 8.17 (2H, m), 8.12 (2H, m), 7.89 (4H, m), 7.75 (3H, m) Met.

Example 19 Stimulation of Platelet Levels Another unexpected property of the disclosed compounds, such as a resonance modifying agent such as A-007, is the ability to increase platelet production in a subject administered a drug. For example, it has been found that subjects undergoing A-007 have enlarged spleen and increased platelet production. This increase in platelet production is particularly advantageous for subjects suffering from thrombocytopenia, eg, from immune or neoplastic diseases, or cytotoxic drugs used to treat such diseases. For example, A-007, or a resonance modifier, or other compound disclosed herein that can promote platelet production is administered to a subject in need of such stimulation. The drug is administered in any manner disclosed herein.

  Mice receiving A-007 according to the disclosed method have been shown to show enlarged spleen (splenomegaly) and increased platelet production. Accordingly, the compounds disclosed herein can result in the production of megakaryocytes and platelets outside the bone marrow (outside the bone marrow).

Example 20 Treatment of Body Surface Lesions The agents disclosed herein can be used to treat a wide variety of body surface lesions such as epilepsy, HSV herpes, psoriasis, and skin cancer. These lesions are treated by applying the drug locally to the lesion for a period sufficient to induce the disappearance or regression of the lesion. For example, the drug is applied to the lesion surface in a 0.25% gel at a dose of 2 g per day for 3-10 days. If necessary, treatment can continue until lesion resolution is achieved.

  Other treatment plans are also available. For example, higher doses or lower doses of active ingredients can be used clinically adequately. The therapeutically effective amount of the drug is determined by the clinician depending on the clinical condition. For example, a total amount of 1-10 g, 2-5 g, or 2-3 g is administered topically during the treatment regimen. Dosage / dose can be once a day (eg, every night), twice a day, or more frequently. The total duration of treatment depends on the clinical condition (eg every day for 3-10 days, eg every day for 5 days) and the treatment plan can be repeated at intervals as needed. For example, treatment can be performed daily for 5 days, followed by no treatment for 25 days, followed by another series of daily treatments for 5 days.

  While the essence of the invention has been illustrated and described in several embodiments, it will be apparent to those skilled in the art that the invention can be modified in detail without departing from the spirit thereof. Claim all modifications that fall within the spirit and scope of the following claims.

FIG. 1 is a schematic representation of the interaction between a conjugated aryl hydrazone and a CD45 + T lymphocyte surface receptor. FIG. 2 shows the structural formula of A-007. FIG. 3 is a schematic diagram of the resonance structure of a conjugated aryl hydrazone.

Claims (148)

It is a compound other than A-007 having the following structural formula,

Where Ar in the formula includes an aryl group substituted with one or more electron withdrawing groups,
X is NH, CH ₂ , O, —, SO ₂ , or B,
Q is-(CH = CH) _m -CH-,
Y and Y ′ are independently selected from N, CH and B;
n is 0 or 1,
m is 0 to 6,
R ¹ is H, a lower alkyl group, an optionally substituted phenyl group or the following structure:

Where R ⁴ is an optionally substituted phenyl group,
R ² is an optionally substituted lower alkyl group, an aralkyl group, a thioalkyl group, an amidoalkyl group, an amino group, an aminoalkyl group, a nitro group, an azido group, and a 4- to 7-membered ring optionally substituted with Z Form a carbocyclic or heterocyclic ring or form an optionally substituted phenyl group with R ³ ;
R ³ together with R ² forms an optionally substituted phenyl group, or is an optionally substituted phenyl group, and
Z is H, a lower alkyl group, a substituted alkyl group, a thioalkyl group, a nitrile group, an amino group, or an optionally substituted 4-7 membered carbocyclic or heterocyclic ring with R ² ,
The compound characterized by the above-mentioned. When X is NH and Y is N, n is 1, R ¹ is R ⁵ or both,
The compound of claim 1, wherein R ⁵ is H or a lower alkyl group.   2. The compound of claim 1, wherein Ar is an aryl group substituted with one or more halogen, haloalkyl group, acyl group, carboxyl group, carboxyamide group, carboalkoxy group, sulfonic acid, cyano group or nitro group. The compound of claim 1, wherein:   The compound of claim 1, wherein Ar contains two or more electron withdrawing groups.   A compound according to claim 4, characterized in that Ar is a dinitrophenyl group. R ¹ is -OR ⁵ , -SR ⁵ , -SO ₂ N (R ⁵ ) ₂ , -SO ₂ CN, -SO ₂ R ⁵ , -SO ₂ N ₃ , N ₃ , -CN, -N (R ⁵ A phenyl group substituted with one or more groups selected from C (O) R ⁶ , -R ⁶ , or combinations thereof;
Each R ⁵ is independently H, a lower alkyl group, a sugar, a fatty acid, an amino acid, a peptide, a nucleoside, or a nucleic acid, and
R ⁶ is a lower alkyl group or a C-glycoside,
The compound of claim 1, characterized in that The compound of claim 1, wherein R ² together with R ³ forms an optionally substituted phenyl group. The compound of claim 1, wherein R ³ is -OR ⁵ , -SR ⁵ , -SO ₂ N (R ⁵ ) ₂ , -SO ₂ CN, -SO ₂ R ⁵ , -SO ₂ N ₃ , a phenyl group substituted by one or more groups selected from N ₃ , —CN, —N (R ⁵ ) C (O) R ⁶ , —R ⁶ , or a combination thereof. The compound of claim 1.   2. The compound of claim 1, wherein n is 0.   2. The compound of claim 1, wherein m is 1, 2, or 3.   2. The compound of claim 1, wherein m is 0 or 1.   The compound of claim 1, wherein both n and m are 1.   2. The compound of claim 1, wherein n is 1 and m is 0. A compound of the following structural formula:

Here, R ⁵ and R ⁶ in the formula are independently -OR ⁷ , -SR ⁷ , -SO ₂ N (R ⁷ ) ₂ , -SO ₂ CN, -SO ₂ R ⁸ , -SO ₂ N ₃ , N ₃ , -CN, -N (R ⁷ ) C (O) R ⁸ , -R ⁷
R ⁷ is H or a lower alkyl group, and
R ⁸ is a lower alkyl group,
14. The compound of claim 13, characterized in that 15. A compound of claim 14 having the structure:

X is CH ₂ , O, SO ₂ , B,
n is 0,
R ¹ is an optionally substituted phenyl group or the following structure:

R ⁴ in the formula is an optionally substituted phenyl group,
R ² is an optionally substituted lower alkyl group, or forms an optionally substituted 4-7 membered carbocyclic or heterocyclic ring with Z, or an optionally substituted phenyl group with R ⁴ Forming, and
R ³ is an optionally substituted phenyl group,
The compound of claim 1, characterized in that R ¹ is -OR ⁵ , -SR ⁵ , -SO ₂ NH ₂ , -SO ₂ CN, -SO ₂ R ⁵ , -SO ₂ N ₃ , N ₃ , -CN, -N (R ⁵ ) C (O 17. The compound of claim 16, which is a phenyl group substituted with one or more groups selected from: R ⁶ , —R ⁶ , or combinations thereof. The compound of claim 16, wherein R ² and R ³ form a phenyl group. Having the following structure:

Here, R ⁷ and R ⁸ in the formula are independently -OR ⁵ , -SR ⁵ , -SO ₂ N (R ⁵ ) ₂ , -SO ₂ CN, -SO ₂ R ⁶ , -SO ₂ N ₃ , N ₃ , -CN, -N (R ⁵ ) C (O) R ⁶ , -R ⁶ ,
Each R ⁵ is independently H or a lower alkyl group, and
R ⁶ is a lower alkyl group,
19. A compound according to claim 18, characterized in that 20. A compound of claim 19 having the structure:

The compound of claim 20 R ⁵ and R ⁶ is -OH. The compound of claim 21, wherein X is CH ₂ and Y is N or CH. 23. The compound of claim 22 having the structure:

Here, R ⁵ in the formula is H, -OR ⁶ , -SR ⁶ , -SO ₂ N (R ⁶ ) ₂ , -SO ₂ CN, -SO ₂ R ⁷ , -SO ₂ N ₃ , N ₃ , -CN , -N (R ⁶ ) C (O) R ⁷ , or -R ⁷ ,
R ¹ and R ⁶ are independently H or a lower alkyl group, and
R ² and R ⁷ are independently a lower alkyl group,
23. The compound of claim 22, characterized in that   24. The compound of claim 23, wherein Ar comprises a nitrophenyl group.   25. The compound of claim 24, wherein Ar comprises a dinitrophenyl group. 26. The compound of claim 25 having the structure:

27. The compound of claim 26, wherein R < ¹ > is H. The compound of claim 27 R ² is a methyl group or an ethyl group. The compound of claim 28 R ⁵ is representing -OR ^6. 30. The compound of claim 29 having the structure:

  A pharmaceutical composition comprising the compound of claim 1 and a pharmaceutically acceptable carrier. 32. The composition of claim 31, wherein the compound has the structure:

  32. The composition of claim 31, wherein the composition comprises a topical gel or solution.   34. The composition of claim 33, wherein the topical gel or solution comprises about 0.25% or more of the compound.   32. The composition of claim 31, wherein the composition comprises a sustained release dosage form.   36. The composition of claim 35, wherein the sustained release dosage form comprises tablets or crystals.   32. The composition of claim 31, wherein the composition is suitable for transport via inhalation.   38. The composition of claim 37, wherein the composition comprises an aerosol.   A pharmaceutical composition comprising a compound of claim 6 and a pharmaceutically acceptable carrier.   A pharmaceutical composition comprising a compound of claim 16 and a pharmaceutically acceptable carrier. A method of stimulating an immune response in a subject in need of immune stimulation,
Selection of subjects in need of immune stimulation;
Administering to said subject in need of immune stimulation a therapeutically effective amount of a resonance modifying compound other than A-007 having the structure:

Where Ar in the formula includes an aryl group substituted with one or more electron withdrawing groups,
X is NH, CH ₂ , O, —, SO ₂ , or B,
Q is-(CH = CH) _m -CH-,
Y and Y ′ are independently selected from N, CH and B;
n is from 0 or 1,
m is 0 to 6,
R ¹ is H, a lower alkyl group, an optionally substituted phenyl group or the following structure:

Where R ⁴ is an optionally substituted phenyl group,
R ² is an optionally substituted lower alkyl group, an aralkyl group, a thioalkyl group, an amidoalkyl group, an amino group, an aminoalkyl group, a nitro group, an azido group, and a 4- to 7-membered ring optionally substituted with Z Form a carbocyclic or heterocyclic ring or form an optionally substituted phenyl group with R ³ ;
R ³ together with R ² forms an optionally substituted phenyl group, or is an optionally substituted phenyl group, and
Z is H, a lower alkyl group, a substituted alkyl group, a thioalkyl group, a nitrile group, an amino group, or an optionally substituted 4-7 membered carbocyclic or heterocyclic ring with R ³ ,
A method involving that.   42. The method of claim 41, wherein selecting the subject in need of immune stimulation comprises selecting a subject with a laboratory value of immunodeficiency.   42. The method of claim 41, wherein selecting the subject comprises selecting a subject infected with an infectious pathogen.   44. The method of claim 43, wherein the infectious pathogen is a virus.   45. The method of claim 44, wherein the pathogen is a human papilloma virus.   42. The method of claim 41, wherein selecting the subject comprises selecting a subject suffering from anal or cervical dysplasia.   49. The method of claim 46, wherein the subject is a subject suffering from dysplasia caused by papillomavirus infection.   42. The method of claim 41, wherein administering the compound comprises administering the resonance modifying compound intravaginally or intraanally to the subject.   49. The method of claim 48, wherein administering the compound comprises introducing a sustained release dosage form intravaginally or intraanally.   52. The method of claim 49, wherein the sustained release dosage form comprises tablets or crystals.   42. The method of claim 41, wherein administering the compound comprises topically administering the compound to a site other than a skin lesion.   Administration of the compound includes topical administration of the compound to a skin surface site, wherein the skin surface is substantially impermeable to the compound and further the immune response is remote from the skin surface site. 42. The method of claim 41, wherein the method is stimulated.   52. The method of claim 51, wherein administration of said compound comprises inhalation administration of said compound.   54. The method of claim 53, wherein the compound is administered by inhalation to a subject suffering from respiratory epithelial infection or tumor.   42. The method of claim 41, wherein the immune response is stimulated in a lymphatic network.   42. The method of claim 41, wherein the compound is applied to a hyperproliferative body surface lesion.   57. The method of claim 56, wherein the hyperproliferative body surface lesion is an anus, cervix, oral cavity, penis, vaginal or vulva lesion.   57. The method of claim 56, wherein the hyperproliferative body surface lesion is a precancerous lesion.   59. The precancerous lesion comprises actinic keratosis, Bowen-like actinic keratosis, arsenic keratosis, Bowen's disease, viral keratosis, leukoplakia, Kelar erythroderma, nevus or hemorrhoids. the method of.   The lesion is a primary neoplastic lesion selected from anal cancer, cervical cancer, basal cell carcinoma or squamous cell carcinoma, melanoma, penile cancer, vulva cancer, vaginal cancer, or oral mucosal cancer. Item 56. The method according to Item 56.   57. The method of claim 56, wherein the body surface lesion is a psoriatic lesion.   42. The method of claim 41, wherein Ar is an aryl group substituted with one or more halogen, haloalkyl group, acyl group, carboxyl group, carboxyamide group, carboalkoxy group, sulfonic acid, cyano group or nitro group.   63. The method of claim 62, wherein the resonance modifying compound is nitrophenyl hydrazone.   63. The method of claim 62, wherein the resonance modifying compound is dinitrophenyl hydrazone.   65. The method of claim 64, wherein the resonance modifying compound is 2,4-dinitrophenylhydrazone.   42. The method of claim 41, wherein Ar comprises a polycyclic group.   42. The method of claim 41, wherein Ar comprises a substituted phenyl group.   42. The method of claim 41, wherein Ar comprises two or more electron withdrawing substituents.   69. The method of claim 68, wherein Ar is a dinitrophenyl group.   42. The method of claim 41, wherein n is 0.   42. The method of claim 41, wherein m is 1, 2, or 3.   42. The method of claim 41, wherein m is 0 or 1.   42. The method of claim 41, wherein both n and m are 1.   42. The method of claim 41, wherein n is 1 and m is 0. 42. The method of claim 41, wherein the resonance modifying compound has the following structure:

Here, R ⁵ and R ⁶ in the formula are independently -OR ⁷ , -SR ⁷ , -SO ₂ N (R ⁷ ) ₂ , -SO ₂ CN, -SO ₂ R ⁸ , -SO ₂ N ₃ , N ₃ , -CN, -N (R ⁷ ) C (O) R ⁸ , -R ⁷
R ⁷ is H or a lower alkyl group, and
R ⁸ is a lower alkyl group,
42. The method of claim 41, wherein: 42. The method of claim 41, wherein the resonance modifying compound has the structure:

The resonance modifying compound comprises the following structure:

Where X in the formula is a dinitrophenyl group,
R ¹ is H, a lower alkyl group, an optionally substituted phenyl group or the following structure:

Wherein R ² is a lower alkyl group or an optionally substituted phenyl group,
R ³ is an optionally substituted lower alkyl group, thioalkyl group, amidoalkyl group, amino group, aminoalkyl group, nitro group, azido group, and a 4-7 membered carbocyclic ring optionally substituted with Z or Forming a heterocycle or optionally forming a phenyl group optionally substituted with R ⁴ ,
R ⁴ together with R ³ forms an optionally substituted phenyl group or is an optionally substituted phenyl group,
42. The method of claim 41, wherein: Method analogous claim 77 R ¹ is H. The resonance modifying compound has the following structure:

Here, R ² in the formula is a lower alkyl group,
R ⁵ is H, OH, -OR ⁶ , -SO ₂ NH ₂ , N ₃ , -CN, -SO ₂ CN, -SO ₂ N ₃ or a lower alkyl group, and
R ⁶ is a lower alkyl group,
79. A method according to claim 78, wherein:   42. The method of claim 41, wherein the administration of the compound comprises applying the compound to the epithelial surface of the subject.   81. The method of claim 80, wherein applying the compound to the epithelial surface of the subject comprises applying the compound to the surface of the skin.   81. The method of claim 80, wherein applying the compound to the epithelial surface of the subject comprises introducing the compound to the skin.   42. The method of claim 41, wherein the method of stimulating an immune response comprises a method of treating a tumor by stimulating the immune response of the subject.   82. The method of claim 81, wherein applying the compound to the skin of the subject comprises applying the compound to an upper or near skin portion of a tumor to stimulate the immune response of the subject.   85. The method of claim 84, wherein the tumor is a malignant tumor.   86. The method of claim 85, wherein the tumor is metastatic cancer.   42. The method of claim 41, further comprising enhancing immune stimulation by exposing the compound to an electromagnetic field that induces an increase in resonance modification of the compound.   42. The method of claim 41, wherein the electromagnetic field is created by a magnetic probe.   90. The method of claim 88, wherein the electromagnetic field is created by a current flowing between two electrodes.   42. The method of claim 41, further comprising monitoring the amplitude of a waveform produced by resonance modification of the compound to determine a response of the subject's immune system to the compound.   94. The method of claim 90, further comprising exposing the compound to an electromagnetic field to induce an increase in resonance modification of the compound when the amplitude is below a therapeutically preferred amplitude.   The method of stimulating an immune response comprises a method of stimulating an immune response by a cell expressing an outer membrane bound protein tyrosine phosphatase or against a cell expressing an outer membrane bound protein tyrosine phosphatase, 42. The method of claim 41.   94. The method of claim 92, wherein the cell is a T lymphocyte and the method of stimulating the immune response comprises stimulating the immune response to an antigen by the T lymphocyte.   94. The method of claim 93, wherein the cell is a cell infected with a CD45 + virus and the method of stimulating the immune response comprises stimulating the immune response against the cell infected with the virus.   95. The method of claim 94, wherein the virus is a papilloma virus or a retrovirus.   96. The method of claim 95, wherein the virus is a papilloma virus.   99. The method of claim 96, wherein the virus is a human papilloma virus.   95. The method of claim 94, wherein the virus is a retrovirus.   99. The method of claim 98, wherein the virus is a human immunodeficiency virus.   The cell is a cell infected with a virus containing an active outer membrane-bound protein tyrosine phosphatase, and the method of stimulating the immune response stimulates the immune response against the cell infected with the virus. 94. The method of claim 92, comprising: A method of monitoring a subject's immune response,
A method comprising: placing a resonance modifying compound in contact with the subject and generating a unique electromagnetic waveform characteristic of the resonance modifying compound other than A-007 having the structure:

Where Ar in the formula includes an aryl group substituted with one or more electron withdrawing groups,
X is NH, CH ₂ , O, —, SO ₂ , or B,
Q is-(CH = CH) _m -CH-,
Y and Y ′ are independently selected from N, CH and B;
n is 0 or 1,
m is 0 to 6,
R ¹ is H, a lower alkyl group, an optionally substituted phenyl group or the following structure:

Where R ⁴ is an optionally substituted phenyl group,
R ² is an optionally substituted lower alkyl group, an aralkyl group, a thioalkyl group, an amidoalkyl group, an amino group, an aminoalkyl group, a nitro group, an azido group, and a 4- to 7-membered ring optionally substituted with Z Form a carbocyclic or heterocyclic ring or form an optionally substituted phenyl group with R ³ ;
R ³ together with R ² forms an optionally substituted phenyl group or is an optionally substituted phenyl group;
Z is H, a lower alkyl group, a substituted alkyl group, a thioalkyl group, a nitryl group, an amino group, or an optionally substituted 4-7 membered carbocyclic or heterocyclic ring with R ^2. , When X is NH and Y is N, n is 1, R ¹ is R ⁵ or both, and
R ⁵ is H or a lower alkyl group, wherein the electromagnetic waveform represents the immune status of the subject, and
Monitoring the waveform produced by said compound for a waveform associated with altered immune function.   102. The method of claim 101, wherein monitoring the waveform comprises detecting a voltage change over time across the compound without introducing an external current or potential difference.   102. The method of claim 101, wherein monitoring of the waveform associated with altered immune function comprises detecting a waveform with a reduced amplitude. A method of treating a cancerous lesion or a precancerous lesion, comprising applying an effective amount of a compound containing a compound other than A-007 having the following structure to the lesion:

Where Ar in the formula includes an aryl group substituted with one or more electron withdrawing groups,
X is NH, CH ₂ , O, —, SO ₂ , or B,
Q is-(CH = CH) _m -CH-,
Y and Y ′ are independently selected from N, CH and B;
n is 0 or 1,
m is 0 to 6,
R ¹ is H, a lower alkyl group, an optionally substituted phenyl group or the following structure:

Where R ⁴ is an optionally substituted phenyl group,
R ² is an optionally substituted lower alkyl group, an aralkyl group, a thioalkyl group, an amidoalkyl group, an amino group, an aminoalkyl group, a nitro group, an azido group, and a 4- to 7-membered ring optionally substituted with Z Form a carbocyclic or heterocyclic ring or form an optionally substituted phenyl group with R ³ ;
R ³ together with R ² forms an optionally substituted phenyl group or is an optionally substituted phenyl group;
Z is H, a lower alkyl group, a substituted alkyl group, a thioalkyl group, a nitryl group, an amino group, or an optionally substituted 4-7 membered carbocyclic or heterocyclic ring with R ^2. , When X is NH and Y is N, n is 1, R ¹ is R ⁵ or both, and
R ⁵ is H or a lower alkyl group,
A method involving that.   105. The method of claim 104, wherein the lesion is a hyperproliferative body surface lesion.   105. The method of claim 104, wherein the hyperproliferative body surface lesion is a precancerous lesion.   106. The method of claim 105, wherein the lesion is an anus, cervix, oral cavity, penis, vagina or vulva.   107. The precancerous lesion comprises actinic keratosis, Bowen-like actinic keratosis, arsenic keratosis, Bowen's disease, viral keratosis, leukoplakia, Kelar erythroderma, nevi or vagina. the method of.   107. The method of claim 106, wherein the precancerous lesion is a papillomavirus-related lesion.   105. The method of claim 104, wherein the lesion is a primary neoplastic lesion.   111. The method of claim 110, wherein the neoplastic lesion is anal cancer, cervical cancer, basal cell or squamous cell carcinoma, melanoma, penile cancer, vulva cancer, vaginal cancer or oral mucosal cancer.   111. The method of claim 110, wherein the primary neoplastic lesion comprises a metastatic neoplastic lesion.   111. The method of claim 110, wherein the lesion is a penile lesion.   111. The method of claim 110, wherein the lesion is a vulva or vaginal lesion.   106. The method of claim 105, wherein the hyperproliferative body surface lesion is a body surface lesion. A method of collecting lymphocytes from a biological tissue comprising exposing the biological tissue to an effective amount of a compound other than A-007 having the structure:

Where Ar in the formula includes an aryl group substituted with one or more electron withdrawing groups,
X is NH, CH ₂ , O, —, SO ₂ , or B,
Q is-(CH = CH) _m -CH-,
Y and Y ′ are independently selected from N, CH and B;
n is 0 or 1,
m is 0 to 6,
R ¹ is H, a lower alkyl group, an optionally substituted phenyl group or the following structure:

Where R ⁴ is an optionally substituted phenyl group,
R ² is an optionally substituted lower alkyl group, an aralkyl group, a thioalkyl group, an amidoalkyl group, an amino group, an aminoalkyl group, a nitro group, an azido group, and a 4- to 7-membered ring optionally substituted with Z Form a carbocyclic or heterocyclic ring or form an optionally substituted phenyl group with R ³ ;
R ³ together with R ² forms an optionally substituted phenyl group or is an optionally substituted phenyl group;
Z is H, a lower alkyl group, a substituted alkyl group, a thioalkyl group, a nitryl group, an amino group, or an optionally substituted 4-7 membered carbocyclic or heterocyclic ring with R ^2. , When X is NH and Y is N, n is 1, R ¹ is R ⁵ or both, and
R ⁵ is H or a lower alkyl group,
Including methods.   117. The method of claim 116, wherein exposing a living body to the effective amount of the compound comprises applying the compound to the skin of the living body.   118. The method of claim 117, wherein application of the compound to the skin comprises applying a topical formulation of the compound to the surface of the skin. A method of treating an infection in a subject, comprising treating the subject with a compound other than A-007 having the following structure, which interacts with a cellular PTP extracellular receptor to activate the receptor: Administering an effective amount,

Where Ar in the formula includes an aryl group substituted with one or more electron withdrawing groups,
X is NH, CH ₂ , O, —, SO ₂ , or B,
Q is-(CH = CH) _m -CH-,
Y and Y ′ are independently selected from N, CH and B;
n is 0 or 1,
m is 0 to 6,
R ¹ is H, a lower alkyl group, an optionally substituted phenyl group or the following structure:

Where R ⁴ is an optionally substituted phenyl group,
R ² is an optionally substituted lower alkyl group, an aralkyl group, a thioalkyl group, an amidoalkyl group, an amino group, an aminoalkyl group, a nitro group, an azido group, and a 4- to 7-membered ring optionally substituted with Z Form a carbocyclic or heterocyclic ring or form an optionally substituted phenyl group with R ³ ;
R ³ together with R ² forms an optionally substituted phenyl group or is an optionally substituted phenyl group;
Z is H, a lower alkyl group, a substituted alkyl group, a thioalkyl group, a nitryl group, an amino group, or an optionally substituted 4-7 membered carbocyclic or heterocyclic ring with R ^2. , When X is NH and Y is N, n is 1, R ¹ is R ⁵ or both, and
R ⁵ is H or a lower alkyl group,
A method involving that.   120. The method of claim 119, further comprising applying an external electromagnetic field to the compound to increase the resonance modification of the compound, thereby increasing the immunostimulatory effect of the compound to treat the infection.   120. The method of claim 119, wherein said infection is a viral infection.   122. The method of claim 121, wherein the virus is a papillomavirus or a retrovirus.   123. The method of claim 122, wherein the virus is a papilloma virus.   124. The method of claim 123, wherein the virus is a human papilloma virus.   123. The method of claim 122, wherein the virus is a retrovirus.   126. The method of claim 125, wherein the virus is a human immunodeficiency virus.   120. The method of claim 119, wherein said infection is a bacterial infection.   120. The method of claim 119, wherein said cell is a T-lymphocyte.   129. The method of claim 128, wherein said cell is a CD45 + lymphocyte.   129. The method of claim 129, wherein said cell is a CD45 + pathogen.   131. The method of claim 130, wherein the pathogen is a human papilloma virus.   120. The method of claim 119, further comprising enhancing the interaction between the compound and the receptor by exposing the compound and cells to ultraviolet light.   120. The method of claim 119, wherein administering the compound to the subject comprises applying the compound to a dysplastic or metaplastic epithelium.   134. The method of claim 133, wherein said epithelium is urogenital epithelium.   135. The method of claim 134, wherein said urogenital epithelium is the vagina, cervix, or anal epithelium.   134. The method of claim 133, wherein administering the compound to the subject comprises applying the compound to squamous cell carcinoma.   137. The method of claim 136, wherein the tumor is anal squamous cell carcinoma.   120. The method of claim 119, wherein administering comprises applying a topical gel.   140. The method of claim 138, wherein said topical gel comprises about 0.25% of said compound.   138. The method of claim 138, wherein applying the compound to the subject comprises applying the compound at least daily for a time sufficient to produce a therapeutic effect.   142. The method of claim 141, wherein administering the compound comprises administering the compound at least daily for 5 days or more.   120. The method of claim 119, wherein the therapeutically effective amount of the compound is 2 grams or more of topical gel per day, and the gel comprises about 0.25% or more of the compound. A method of activating a cellular protein tyrosine phosphatase present in a cell membrane, wherein the cell is treated with a compound other than A-007 having the following structure and a sufficient amount of a resonance modifying compound to activate the protein tyrosine phosphatase Including exposure to

Where Ar in the formula includes an aryl group substituted with one or more electron withdrawing groups,
X is NH, CH ₂ , O, —, SO ₂ , or B,
Q is-(CH = CH) _m -CH-,
Y and Y ′ are independently selected from N, CH and B;
n is 0 or 1,
m is 0 to 6,
R ¹ is H, a lower alkyl group, an optionally substituted phenyl group or the following structure:

Where R ⁴ is an optionally substituted phenyl group,
R ² is an optionally substituted lower alkyl group, an aralkyl group, a thioalkyl group, an amidoalkyl group, an amino group, an aminoalkyl group, a nitro group, an azido group, and a 4- to 7-membered ring optionally substituted with Z Form a carbocyclic or heterocyclic ring or form an optionally substituted phenyl group with R ³ ;
R ³ together with R ² forms an optionally substituted phenyl group or is an optionally substituted phenyl group;
Z is H, a lower alkyl group, a substituted alkyl group, a thioalkyl group, a nitrile group, an amino group, or an optionally substituted 4-7 membered carbocyclic or heterocyclic ring with R ² ,
A method involving that.   145. The method of claim 143, wherein exposing the cell to the amount of the resonance modifying agent comprises applying the resonance modifying agent locally to or in the vicinity of the cell. A method of stimulating an immune response in a subject in need of immune stimulation,
Selection of subjects in need of immune stimulation;
Administering to said subject a therapeutically effective amount of a compound other than A-007 having the structure:

Where Ar in the formula includes an aryl group substituted with one or more electron withdrawing groups,
X is NH, CH ₂ , O, —, SO ₂ , or B,
Q is-(CH = CH) _m -CH-,
Y and Y ′ are independently selected from N, CH and B;
n is from 0 or 1,
m is 0 to 6,
R ¹ is H, a lower alkyl group, an optionally substituted phenyl group or the following structure:

Where R ⁴ is an optionally substituted phenyl group,
R ² is an optionally substituted lower alkyl group, an aralkyl group, a thioalkyl group, an amidoalkyl group, an amino group, an aminoalkyl group, a nitro group, an azido group, and a 4- to 7-membered ring optionally substituted with Z Form a carbocyclic or heterocyclic ring or form an optionally substituted phenyl group with R ³ ;
R ³ together with R ² forms an optionally substituted phenyl group, or is an optionally substituted phenyl group, and
Z is H, a lower alkyl group, a substituted alkyl group, a thioalkyl group, a nitrile group, an amino group, or an optionally substituted 4-7 membered carbocyclic or heterocyclic ring with R ² ,
A method involving that. A method for treating a subject suffering from a hyperproliferative disease,
Selection of subjects in need of immune stimulation;
Administering to said subject a therapeutically effective amount of a cytotoxic compound of the structure:

Where Ar in the formula includes an aryl group substituted with one or more electron withdrawing groups,
X is NH, CH ₂ , O, —, SO ₂ , or B,
Q is-(CH = CH) _m -CH-,
Y and Y ′ are independently selected from N, CH and B;
n is from 0 or 1,
m is 0 to 6,
R ¹ is H, a lower alkyl group, an optionally substituted phenyl group or the following structure:

Where R ⁴ is an optionally substituted phenyl group,
R ² is an optionally substituted lower alkyl group, an aralkyl group, a thioalkyl group, an amidoalkyl group, an amino group, an aminoalkyl group, a nitro group, an azido group, and a 4- to 7-membered ring optionally substituted with Z Form a carbocyclic or heterocyclic ring or form an optionally substituted phenyl group with R ³ ;
R ³ together with R ² forms an optionally substituted phenyl group, or is an optionally substituted phenyl group, and
Z is H, a lower alkyl group, a substituted alkyl group, a thioalkyl group, a nitrile group, an amino group, or an optionally substituted 4-7 membered carbocyclic or heterocyclic ring with R ² ,
A method involving that. A method for treating thrombocytopenia,
Selection of subjects suffering from thrombocytopenia,
Administering to the subject a therapeutically effective amount of a cytotoxic compound other than A-007 having the structure:

Where Ar in the formula includes an aryl group substituted with one or more electron withdrawing groups,
X is NH, CH ₂ , O, —, SO ₂ , or B,
Q is-(CH = CH) _m -CH-,
Y and Y ′ are independently selected from N, CH and B;
n is from 0 or 1,
m is 0 to 6,
R ¹ is H, a lower alkyl group, an optionally substituted phenyl group or the following structure:

Where R ⁴ is an optionally substituted phenyl group,
R ² is an optionally substituted lower alkyl group, an aralkyl group, a thioalkyl group, an amidoalkyl group, an amino group, an aminoalkyl group, a nitro group, an azido group, and a 4- to 7-membered ring optionally substituted with Z Form a carbocyclic or heterocyclic ring or form an optionally substituted phenyl group with R ³ ;
R ³ together with R ² forms an optionally substituted phenyl group, or is an optionally substituted phenyl group, and
Z is H, a lower alkyl group, a substituted alkyl group, a thioalkyl group, a nitrile group, an amino group, or an optionally substituted 4-7 membered carbocyclic or heterocyclic ring with R ² ,
A method involving that. 148. The method of claim 147, wherein the compound is a compound having the structure:

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