Heptamethine dyes

IR-808, A heptamethine dye^[1]

Names
IUPAC name Br 2-[(E)-2-[(3E)-2-chloro-3-\\{2-[(2E)-3,3-dimethyl-1-pentyl-2,3-dihydro-1H-indol-2-ylidene]ethylidene}cyclohex-1-en-1-yl]ethenyl]-3,3-dimethyl-1-pentyl-3H-indol-1-ium
Other names Otterblue
Properties
Chemical formula	C40H52Cl1N2
Molar mass	596.307 g/mol
Appearance	Black Powder
Except where otherwise noted, data are given for materials in their standard state (at 25 °C [77 °F], 100 kPa).
Infobox references

Heptamethine dyes are a subclass of chemical compounds within the indocyanine dye family and are currently being explored for their unique uses in medical diagnostics, the development of theranostics, the individualized treatment of cancerous patients with the aid of PDT, co-administration with other drugs, and irradiation.^[2][3] In tests it is administered intravenously and, depending on hepatic functionality, have been found to be nontoxic at doses of 500 times that needed to fluoresce the tumor cells.^[4] The therapeutic properties appear safe and viable due to the light wavelength absorption being just outside the visible spectrum in the Near Infra-Red range.

The characteristics of heptamethine dyes can be understood by comparing research data on different dyes within the family. For example, the IR-780 dye exhibits a peak spectral absorption of around 780 nm depending on the diluting agent.^[4] Tests have also identified the viability of other analogs including IR-783 and MHI-148.^[4] One specific analog of interest is IR-808 which has shown to exhibit not only similar binding and illumination characteristics of the IR-780 analog, but it has shown to have significant photodependent cytotoxicity in various myeloma cell lines and moderate cytotoxicity in HeLa cells both in vivo and in vitro without the need to conjugate and likely change the conformation ability of co-administered cancer drugs such as paclitaxel.^[2]

Optical properties

The absorption and fluorescence spectrum of heptamethine dyes are in the near infrared region and vary largely on the solvent and concentration.^[5] IR-808 absorbs mainly between 775 nm and 796 nm and emits fluorescence between 808 nm and 827 nm with a large red shift as expected in the serum sample.^[2] These absorption rates are important for their medical applications due to the image capturing capabilities. Dyes with good pharmacokinetics and fluorescence above 680 nm are considered viable in biomedical imaging due to their significant tissue penetration and imaging signature relative to background frequencies.^[6] IR-808 and other analogs produced well defined images with excellent visualization of cancer tumor cells.

Toxicity and side-effects

Heptamethine dyes are metabolized in the liver and excreted via the bile ducts.^[4] In experiments this was exhibited by viewing extensive fluorescence only in the fecal matter of mice without tumors in the removed intestines after the dye had time to propagate in the test subject’s body.^[4] This implies that the dye is effectively removed from the systemic system by the liver and cleared from major organs into the intestinal tract via the bile duct within 48 hours.^[2] Further tests in humans and imaging exposure to NIR light have yet to be performed. When the mice were injected with 500 times the necessary dose there appeared to be no side effects noticed which indicates that this dye contains excellent possibilities as a biomedical imaging tool.^[2]

Uses

Diagnostics

figure 2 - Image of tumor specific fluoresced dye in lab mice overlaid with xray image to locate tumor

The in vivo accumulation of dyes in tumor cells are being explored using HeLa cells in mice to explore the possible use as a tool in the growing field of theranostics.^[3] These tools show a promising ability to increase the survival rate of individuals undergoing cancer treatment by using a personally designed treatment method based on the increasing amount of patient specific diagnostic data available. In heptamethine dyes the contrast index, which measures the relative emission of the dye fluorescence in tumor cells relative to non tumor cells, was found to be considerable if the value observed exceeded 2.5.^[7] In the IR-808 complex the CI exhibits 25 at 10 days after the HeLa tumor producing mice are injected with the dye which indicates this specific complex exceeds the criteria as an effective diagnostic agent.^[2] Upon further investigation with rhodamine 123 the dye was found to target specifically the mitochondria of the cells.^[2] This indicates the dye targets the portion of the tumor cell which is vital to cellular survival.^[8]

While the IR-808 is attached to the cancer cells the target organism can be imaged with NIR fluorescent imaging techniques. When overlaid with the x-ray of the organism the exact location and extent of the tumor cells can be determined (figure 2). Being able to detect small tumors with great accuracy and no side effects is a groundbreaking and very powerful tool in early diagnosis of cancer tumors. It has been shown that physicians using this type of dye improve the accuracy of detecting cancer cells to around 96%.

Perfusion diagnostics of tissues and organs

IR-808 and other analogs have been shown to perform as a marker in the assessment of the perfusion of tissues and organs in HeLa tumor growing mice.^[2] The light used to excite the tumor bound dye is a near infrared light. The fluorescent signature can then captured by a recording device such as the Kodak in-vivo Professional Imaging System.^[4] Being able to capture the image means that the extent and location of cancerous cells in capillary beds throughout the body can be assessed and documented. Therefore, the possible uses in medicine of just the imaging effects of heptamethine dyes would allow the earliest detection and further monitoring of tumor cell growth in one of the smallest transport systems of the body without the use of radioactive chemicals in a very short time.^[8] The capabilities of heptamethine dyes are also novel in that they recently have been found to show the capabilities of killing tumor cells when co-administered with a PDT.^[2]

Uses in PDT - Photodynamic Therapy

The heptamethine dye IR-808 is unique in that it has been found to not only attach to cancer cells independent of other mechanisms, but it has also been found to exhibit cytotoxic properties when irradiated within 6 hours of administration of the dye.^[2] The tumor cells selectively take up the dye as previously described and then are exposed to light wavelengths at the same frequency, in this case 808 nm, as the dye absorbs. In mice the cancer tumor cells of the LLC cell line with the dye and irradiation were found to have significantly decreased in both mass and volume.^[2] HeLa cell viability in vitro appeared to have the lease amount of cytotoxicity when compared to other cell lines and similar Photodynamic Therapy methods.^[2] Further research into the efficacy and methods of varying cell lines would verify the validity of its use as a legitimate anti-tumor agent independent of other tumor suppressing drugs. Nevertheless, the opportunities of a biomedically safe and minimally invasive cancer targeting, imaging, diagnosing, and tumor reducing agent in IR-808 and possibly other heptamethine dyes remain an exciting and promising field worth further research.

Patents

International patents on the use of heptamethine dyes for use in photographic use have also been approved and instituted. Fuji film placed a patent titled “Heptamethine cyanine compound, near infrared absorbing ink, near infrared absorbing sheet and silver halide photographic material” in 1999 and a follow-up international patent 6072059 in 2000.

Heptamethine dyes for near infrared fluorescent imaging are manufactured by such companies as Intrace Medical, Lumiprobe, Cyandye, LI-COR Biosciences - GmbH and others.

References

1. Tan X, Luo S, Wang D, et al. A NIR heptamethine Dye with intrinsic cancer targeting, imaging and photosynthesizing properties. Journal of Biomaterials China. 33-7 (2011), pp. 2230-2239.
2. Tan X, Luo S, Wang D, Su Y, Cheng T, Shi C (March 2012). "A NIR heptamethine dye with intrinsic cancer targeting, imaging and photosensitizing properties". Biomaterials. 33 (7): 2230–9. doi:10.1016/j.biomaterials.2011.11.081. PMID 22182749.
3. F. Pene, E. Courtine, A. Cariou, J.P. Mira. Toward theranostics. Crit Care Med, 37 (2009), pp. S50–S58
4. Yang X, Shi C, Tong R, Qian W, Zhau HE, Wang R, Zhu G, Cheng J, Yang VW, Cheng T, Henary M, Strekowski L, Chung LW (May 2010). "Near IR heptamethine cyanine dye-mediated cancer imaging". Clin. Cancer Res. 16 (10): 2833–44. doi:10.1158/1078-0432.CCR-10-0059. PMC 2871283. PMID 20410058.
5. Goldsmith SJ (April 1997). "Receptor imaging: competitive or complementary to antibody imaging?". Semin Nucl Med. 27 (2): 85–93. PMID 9144853.
6. "Near Infrared Dyes - Cytodiagnostics".
7. Andreev OA, Dupuy AD, Segala M, Sandugu S, Serra DA, Chichester CO, Engelman DM, Reshetnyak YK (May 2007). "Mechanism and uses of a membrane peptide that targets tumors and other acidic tissues in vivo". Proc. Natl. Acad. Sci. U.S.A. 104 (19): 7893–8. doi:10.1073/pnas.0702439104. PMC 1861852. PMID 17483464.
8. Zhang E, Zhang C, Su Y, Cheng T, Shi C (February 2011). "Newly developed strategies for multifunctional mitochondria-targeted agents in cancer therapy". Drug Discov. Today. 16 (3-4): 140–6. doi:10.1016/j.drudis.2010.12.006. PMID 21182981.